SummaryThe yolk syncytial layer (YSL) has been regarded as one of the main obstacles for a successful cryopreservation of fish embryos. The purpose of this study was to identify and characterize the YSL in Prochilodus lineatus, a fish species found in southeastern Brazil and considered a very important fishery resource. Embryos were obtained through artificial breeding by hormonal induction. After fertilization, the eggs were incubated in vertical incubators with a controlled temperature (28• C). Embryos were collected in several periods of development up to hatching and then fixed with 2% glutaraldehyde and 4% paraformaldehyde in 0.1 M sodium phosphate buffer (pH 7.3). Morphological analyses were carried out under either light, transmission or scanning electron microscopy. The formation of the YSL in P. lineatus embryos starts at the end of the cleavage stage (morula), mainly at the margin of the blastoderm, and develops along the embryo finally covering the entire yolk mass (late gastrula) and producing a distinct intermediate zone between the yolk and the endodermal cells. The YSL was characterized by the presence of microvilli on the contact region with the yolk endoderm. A cytoplasmic mass, full of mitochondria, vacuoles, ribosomes, endomembrane nets and euchromatic nuclei, indicated a high metabolic activity. This layer is shown as an interface between the yolk and the embryo cells that, besides sustaining and separating the yolk, acts as a structure that makes it available for the embryo. The structural analyses identified no possible barriers to cryoprotectant penetration.

The performance of Nile tilapia (Oreochromis niloticus) raised in cages furnished with an automatic dispenser, supplied at different frequencies (once per hour and once every two hours) and periods (daytime, nighttime and both) was evaluated. Eighteen 1.0m 3 cages were placed into a 2000m 2 pond, two meters deep with a 5% water exchange. One hundred and seventy tilapias, with initial weight of 16.0±4.9g, were dispersed into each 1m 3 cage and the feed ration was adjusted every 21 days with biometry. Data was collected from March to July (autumn and winter). Significant difference to final weight (P<0.05) among treatments was observed. The increase in feeding frequency improves the productive performance of Nile tilapias in cages and permitted better management of the food. The better feed conversion rate for high feeding frequency (24 times day -1 ) can result in saving up to 360kg of food for each ton of fish produced, increasing the economic sustenance for tilapia culture and suggesting less environmental pollution.

RESUMO -Este trabalho teve por objetivo aperfeiçoar a técnica de reprodução induzida existente para rã-touro, com o intuito de aumentar a taxa de fecundidade e viabilizar seu uso pelo produtor. As doses hormonais para a indução da ovulação e espermiação seguiram as propostas de FALCON e CULLEY (1995) e ALONSO (1997); entretanto, a técnica de fertilização artificial foi adaptada da metodologia para reprodução artificial de peixes com ovos não-aderentes (WOYNAROVICH e HORVÁTH, 1983). A técnica proposta apresenta as seguintes etapas: I) sincronização da ovulação e da espermiação, por meio de hormônio liberador de gonadotropina ((Des-Gli 10 , DHis(Bzl) 6 , Pro-NHEt 9 )-LHRH)); II) extração dos óvulos de cada fêmea (1 a 2 minutos); III) fertilização dos óvulos (2 minutos) com líquido espermático diluído em 100 mL de água; IV) hidratação dos ovos em 10 a 20 litros de água; e V) incubação dos ovos em quadros de tela de 1x 0,70 m, com malha de 1 mm. As taxas de fertilização obtidas com as modificações propostas foram superiores a 60%. Ressalta-se ainda que a técnica propiciou a obtenção, a partir de um mesmo animal, de várias desovas, sendo que cada fêmea pode ovular em intervalos de, aproximadamente, 45 dias.Palavras-chave: fertilização artificial, LHRHa, Rana catesbeiana, reprodução Spawning Inducing by Analog LHRH and Artificial Fertilization of Bullfrog (Rana catesbeiana)ABSTRACT -The objective of this study was to improve the artificial fertilization techniques for bullfrog, to increase the fertilization rate and become their use accessible for frog producers. Hormone dosages for inducing ovulation and spermiation followed those proposed by FALCON and CULLEY (1995) and ALONSO (1997); however, the artificial fertilization technique was adapted from the available methodology for artificial reproduction of fish with non-adhering eggs (WOYNAROVICH e HORVÁTH, 1983). The proposed technique consists of the following steps: I) synchronization of ovulation and spermiation through gonadotropin releasing hormone ((Des-Gli 10 , D-His(Bzl) 6 , Pro-NHEt 9 )-LHRH)); II) complete ovule extrusion in every female (1 to 2 minutes); III) ovule fertilization (2 minutes) with sperm diluted in 100 mL water; IV) ova hydration in 10 -20 L water; V) ova incubation in screen frames, measuring 1,0 x 0,7 m, with 1 mm screen size. Fertilization rate obtained with the proposed modifications stayed above 60 %. It should also be emphasized that the proposed techniques allowed to obtain several spawns from the same frog, and each female was able to ovulate, approximately, every 45 days. IntroduçãoA reprodução de rãs na maioria dos ranários comerciais é cíclica, ocorrendo picos de desovas no verão e somente algumas na primavera, no outono e inverno. Esta sazonalidade da reprodução compromete o cronograma de produção dos ranários, pois a produção de girinos e imagos fica restrita a alguns meses do ano; conseqüentemente, os galpões de recria e os tanques de girinos permanecem vazios durante vários meses.As fêmeas adultas de rã-touro permanecem madura...

SummaryWhile the freezing techniques of mammal embryos have been providing promising results, the cryopreservation of teleostean eggs and embryos have remained unsuccessful up to now. Therefore, this work aimed to develop a procedure of cryogenic preservation of embryos of Prochilodus lineatus and to observe, at both structural and ultrastructural levels, the morphological alterations that took place after the application of freezing/thawing techniques. The embryos at the morula stage could not tolerate exposure to the cryoprotectants ethylene glycol monomethyl ether, propylene glycol monomethyl ether, methanol, dimethyl sulphoxide and propylene glycol, presenting 100% of mortality. Embryos at the 4-to 6-somites stage tolerated exposure to propylene glycol and dimethyl sulphoxide, and the results revealed no significant differences (α = 0.05) regarding survival from both treatments. None of the freezing, thawing and hydration protocols was effective on preserving embryo viability. The ultrastructural analyses of frozen and thawed embryos showed that cells from ectoderm, somites, notochord and endoderm were structurally intact, with well preserved nuclei and mitochondria. The yolk globules were able to tolerate the freezing process, but the yolk syncytial layer was unorganized, displaying an electron-dense and compacted appearance, collapsed reticules, nuclei with modified chromatin and ruptures on the plasmatic membrane at the contact zone with endoderm. It might be concluded that the procedures tested for freezing were unable to avoid the formation of intracellular ice crystals, leading to drastic morphological modifications and making P. lineatus embryos unviable.

RESUMO -Objetivou-se com a execução deste trabalho avaliar, em nível de campo, o desempenho zootécnico de girinos de rã-touro (Rana catesbeiana) criados em ranários comerciais que utilizam o sistema anfigranja, e simultaneamente estimar os valores de consumo diário de ração para compor uma tabela de referência para a alimentação dos animais. Os resultados encontrados nas unidades de observações monitoradas (aproximadamente 249 mil animais) atingiram os seguintes índices médios: a mortalidade (em %) variou de: 0,7 a 35,2 (média 5,9%); o ganho de peso (g. p/dia): de 0,03 a 0,22 (média 0,11 g/dia); e a conversão alimentar: de 0,92 a 2,75 : 1; (média 1,50 : 1). Comparados com dados preliminares, tais resultados indicam uma significativa otimização dos índices zootécnicos, demonstrando que houve um relativo ganho de produtividade com o aperfeiçoamento das técnicas de manejo implementadas. A redução do valor médio da conversão alimentar aparente foi provocada principalmente pelo adequado uso da tabela de referência, cujos valores foram ratificados durante o trabalho de monitoramento das unidades. Tal tabela possibilita o cálculo da quantidade de alimento a ser oferecido diariamente, baseado nos peso médio dos girinos e na estimativa do percentual de consumo, para temperatura da água entre 18 a 29 graus centígrados, (girinos de 0,1 a 26 gramas de peso médio).Palavras-chave: criação de girinos; índices zootécnicos; rã-touro; ranicultura; Sistema Anfigranja; tabela de alimentação. Performance of Bullfrog Tadpoles (Rana catesbeiana) Raised in the "Amphifarm" System and Feeding TablesABSTRACT -The objective of this work was to evaluate (under field conditions), the performance of bullfrog tadpoles (Rana catesbeiana) raised on commercial frog farms using the "Amphifarm" Systems, and, at the same time, to estimate daily food intake, in order to obtain a reference table for feeding the animals. The results found on the observation units (approximately 249 mil animals) showed the following indexes: mortality(%): range from 0.7 to 35.2, the average being 5.9; weight gain (g/day) varied from 0.03 to 0.22, the average being 0.11; feed conversion ranged from 0.92 to 2.75:1, the average being 1.50:1. Compared to previous data, these results indicate a significant improvement in performance indexes, showing that there was a relative gain in productivity due to the improvement of management techniques. The reduction in average feed conversion was caused mostly by adequate use of the reference table, the values of which were confirmed during a follow-up of the observation units. This table allows the calculation of daily food offer, based on tadpole average weight and on estimated intake as a percentage of body weight, for water temperatures varying from 18 to 29 C and body weight ranging from 0.1 to 26 g.