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In fish, in vitro fertilization is an important reproductive tool used as first step for application of others biotechniques as chromosome and embryo manipulation. In this study, we aimed to optimize gamete quality and their short-term storage from the yellowtail tetra Astyanax altiparanae, for future application in laboratory studies. Working with sperm, we evaluated the effects of spawning inducers (carp pituitary gland and Ovopel® [(D-Ala6, Pro9-NEt) – mGnRH+metoclopramide]) and the presence of female on sperm motility. Additionally, we developed new procedures for short-term storage of sperm and oocytes. Briefly, sperm motility was higher when male fish were treated with carp pituitary gland (73.1±4.0%) or Ovopel® (79.5±5.5%) when compared with the control group treated with 0.9% NaCl (55.6±27.2%; P=0.1598). Maintenance of male fish with an ovulating female fish also improved sperm motility (74.4±7.4%) when compared with untreated male fish (42.1±26.1%; P=0.0018). Storage of sperm was optimized in modified Ringer solution, in which the sperm was kept motile for 18 days at 2.5°C. The addition of antibiotics or oxygen decreased sperm motility, but partial change of supernatant and the combination of those conditions improve storage ability of sperm. Fertilization ability of oocytes decreased significantly after storage for 30, 60 90 and 120 min at 5, 10, 15 and 20°C when compared with fresh oocytes (P=0.0471), but considering only the stored samples, the optimum temperature was 15°C. Those data describe new approaches to improve semen quality and gametes short-term storage in yellowtail tetra A. altiparanae and open new possibilities in vitro fertilization. [ABSTRACT FROM PUBLISHER]

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Interspecific hybrids of fish can offer important advances in the zootechnical exploration of this animal group. However, hybrids can pose serious biological risks to the environment and natural populations, making genetic characterization and monitoring of hybrids produced in fish culture important concerns. In the present study, individuals of the hybrid ‘Piaupara’ were characterized by fluorescence in situ hybridization (FISH) using sex-specific probes to differentiate the respective parental contributions from female Leporinus macrocephalus (Piauçu) and male Leporinus elongatus (Piapara). The application of this technique demonstrated a difference in hybridization patterns between males and females of L. elongatus and L. macrocephalus, mainly due to the fact that the NOR-bearing chromosomes of L. elongatus contain some homology with the probe. This allowed us to detect a clear sex-specific distinction of the hybrid ‘Piaupara’, demonstrating that this technique is well-suited for the characterization of these hybrids. Moreover, this technique offers an important methodological advance for the study of diverse sex chromosome systems of Leporinus, supplying valuable information about the origin and establishment of a recently described sexual system in this genus. Copyright © 2009 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

This study analyzed semen parameters and the ultrastructure of spermatozoa of Brycon vermelha. The semen was white and viscous, with a mean volume of 5.0±2.6 mL/kg body weight and mean spermatozoon concentration of 4.3±0.8×10(10) spermatozoa/mL. The estimated motility rate was 90%, with 50% of spermatozoa motile at 35.0±0.1 s and 100% immotile by 46.5±0.1 s. The spermatozoon of B. vermelha had a distinct head, midpiece, and flagellum. The ovoid head measured 1.9±0.2 μm by 1.3±0.1 μm, with its volume almost completely occupied by the nucleus, and was enveloped by an irregular nuclear membrane, with no acrosome vesicle. The nuclear fossa held the centriole complex and the initial segment of flagellum. The midpiece was symmetrical and measured 1.3±0.3μm. Mitochondria were scarce and restricted to the anterior region, while vesicles were absent. The posterior region of the midpiece was characterized by the absence of mitochondria and the presence of the cytoplasmic sheath. The flagellum, enclosed by the flagellar membrane, measured 29.6±3.4 μm, and possessed an axial filament containing a 9+2 microtubule pattern. The spermatozoa of B. vermelha appeared similar in structure to those of fish that breed through external fertilization, thus classifying them as uniflagellate anacrosomal aquasperm, or Type 1 aquasperm., (Copyright © 2015 Elsevier B.V. All rights reserved.)

To ascertain the origin of B chromosomes in 2 fish species of the genus Prochilodus, i.e. P. lineatus and P. nigricans, we microdissected them and generated B-specific DNA probes. These probes were used to perform chromosome painting in both species and in 3 further ones belonging to the same genus (P. argenteus, P. brevis and P. costatus). Both probes hybridized with the B chromosomes in P. lineatus and P. nigricans, but with none of the chromosomes in the 5 species. This indicates that the B chromosomes have low similarity with DNAs located in the A chromosomes and suggests the possibility that the B chromosomes in the 2 species have a common origin. The most parsimonious explanation would imply intergeneric hybridization in an ancestor of P. lineatus and P. nigricans yielding the B chromosome as a byproduct, which remained in these 2 species after their phylogenetic origin, but was perhaps lost in other Prochilodus species. This hypothesis predicts that B chromosomes are old genomic elements in this genus, and this could be tested once a species from a relative genus would be found showing homology of its A chromosomes with the B-probes employed here, through a comparison of B chromosome DNA sequences with those in the A chromosomes of this other species.