Eduardo Antônio Sanches

Dose inseminante para fertilização artificial de ovócitos de jundiá cinza, Rhamdia quelen (Quoy & Gaimardm, 1824)

Two females with colored urogenital organ, rounded off abdomen, color egg released, and uniform size and four males with semen released under abdomen pressure were selected to determine the best insemination dose for artificial fertilization of grey jundia, Rhamdia quelen (Quoy & Gaimardm, 1824), oocytes. Females and males received two doses of carp pituitary extract: the first one of 0.5 and 0.25 mg.kg-1 and the second of 5.0 and 2.5 mg.kg-1 for females and males, respectively. Both females produced 169 g of eggs with 1,141 oocytes.oocytes g-1. The males produced a total of 23.6 mL of semen. From all males semen pool sperm concentration (1.97 x 1010 spermatozoa.mL-1), survival sperm index (96%) and sperm morphology were analyzed (66.9% of normal spermatozoa). The fertilization assay was conducted as a complete randomized experimental design with five treatments ((6x103, 6x104, 6x105, 6x106, 2x107 spermatozoa.oocyte-1) and four replications. After eight hours of incubation, the fertilization rates for each treatment were analyzed. The best spermatozoa.oocyte-1 rate was of 89.497 spermatozoa.oocyte-1 that produced a maximum fertilization rate of 86.68%.

Artificial fertilization of oocytes and sperm activation in pacu: effects of the spermatozoa:oocyte ratio, water volume, and in natura semen preservation

Objetivou-se foi avaliar a fertilizacao artificial e a duracao da motilidade espermatica em pacus com diferentes doses inseminantes, volumes de agua e preservacao do semen in natura. Foram realizados quatro experimentos para avaliacao do efeito de doses inseminantes (7x103, 7x104, 7x105, 7x106 e 7x107 espermatozoides ovocito-1) sobre a fertilizacao artificial dos ovocitos; do efeito do volume de agua (0,5; 15,0; 30,0; 45,0 e 60,0 mL de agua mL-1 de ovocitos) com doses inseminantes de 105.481 e 210.963 espermatozoides ovocito-1; do efeito de diluicao do semen (0,005; 0,05; 0,5 e 5,0 µL de semen mL-1 de agua) sobre a duracao da motilidade espermatica; e do efeito do armazenamento a 15 oC por 9 h sobre a duracao da motilidade espermatica e o indice de sobrevivencia espermatica. Os maiores resultados obtidos foram: doses inseminantes entre 7x103 e 7x107 espermatozoides ovocito-1; 15 a 60 mL de agua mL-1 de ovocitos; diluicao de 0.005 µL semen mL-1 de agua e 98,65% de sobrevivencia espermatica ate o tempo de preservacao de 2h45min36s. A preservacao a 15oC por 9 horas nao influencia a duracao da motilidade espermatica. As maiores taxas de fertilizacao podem ser observadas no emprego de 0,27 a 270 µL de semen mL-1 de ovocitos, com 15 a 60 mL de agua para ativacao.

Dose inseminante para fertilização artificial de ovócitos de dourado

Objetivou-se determinar a dose inseminante adequada para uso na fertilizacao artificial de ovocitos de dourado (Salminus brasiliensis). Os ovocitos foram distribuidos em delineamento inteiramente casualizado, e fertilizados com uma das relacoes espermatozoides/ovocito 6,0×103; 6,0×104; 6,0×105; 6,0×106 ou 3,0×107, cada uma com quatro repeticoes. Considerou-se unidade experimental uma incubadora de volume util de 2,5 L, contendo 2,0 mL de ovocitos nao-hidratados. As taxas de fertilizacao foram mensuradas 8 horas apos o inicio da fertilizacao. Com intuito de verificar possiveis efeitos da diluicao seminal na movimentacao dos espermatozoides, realizou-se a mensuracao do tempo de duracao da motilidade espermatica dos espermatozoides de dourado, ativados por meio de diferentes relacoes de diluicao: 6,8×10-5; 6,8×10-4; 6,8×10-3; 6,8×10-2; 3,4×10-1 e 1,0 mL de semen por mL de agua. O tempo de duracao da motilidade foi avaliado em delineamento inteiramente casualizado composto de seis tratamentos e tres repeticoes. As taxas de fertilizacao apresentaram relacao quadratica com o numero de espermatozoides por ovocito. As relacoes de diluicao do semen tiveram efeito inversamente proporcional sobre a duracao da motilidade espermatica. A relacao que proporcionou melhores taxas de fertilizacao artificial de ovocitos de dourado (Salminus brasiliensis) foi de 30.722 espermatozoides por ovocio.

Desempenho reprodutivo e zootécnico e deposição de lipídios nos hepatócitos de fêmeas de tilápia-do-nilo alimentadas com rações de diversos níveis energéticos

The experiment was carried out to evaluate the reproductive and animal performance and lipids deposition in the liver tissue of the Nile tilapia female, fed diets containing different levels of digestible energy, obtained by soybean oil inclusion. Three hundred breeding females and 100 breeding males were used, distributed in a completely randomized design with five treatments and four replications. The broods were fed diets containing 35% crude protein and 2,700; 2,950; 3,200; 3,450 and 3,700 kcal energy digestible/kg feed. The animals were stocked in hapas for 93 days. The reproductive and animal performance were not influenced by the energy levels. These results may be associated with the short period of time that the brood were fed the experimental diets. However, increasing energy levels caused a linear increase in lipids inclusions in hepatocytes. Thus, the increasing levels of digestible energy provided to Nile tilapia females, obtained by including soybean oil, only promoted increased lipid deposition in hepatocytes.

Níveis de energia digestível sobre os desempenhos reprodutivo e zootécnico e a deposição de lipídios nos hepatócitos de machos de tilápia-do-nilo

This experiment was carried out to evaluate the reproductive and zootechnical performance, and lipid deposition in the liver tissue of the Nile tilapia males fed diets containing different levels of digestible energy obtained from inclusion of soybean oil. Four hundred 400 broodfish (300 females and 100 males) were assigned to a completely randomized design composed of five levels of digestible energy (2,700, 2,950, 3,200, 3,450 and 3,700 kcal digestible energy.kg of feed-1) and four replications. The breeding were fed diets containing 35% of crude protein and submitted to hapas reproductive management for 101 days. The model adjusted by multiple regression analysis suggests the best result for sperm concentration and percentage of normal sperm for breeding fed diets containing 3.465.56 and 3.443.43 kcal digetible energy.kg of feed-1, producing 7.98 × 109 sperm.mL of semen-1 and 38.98% of normal sperm, respectively. The diet energy levels also caused an increase of lipid inclusion in the hepatocytes. The sperm production, seminal pH, sperm survival index and sperm activation time were not affected by energetic levels of the diets. The zootechnical performance was not affected by the energy levels of the diets either. The energy levels in the diets caused a linear increase in the lipid inclusion in the hepatocytes and affected the sperm quality by stimulating the spermatozoa production and the improving of normality indices of sperm morphology in energy levels around 3,450 kcal digestible energy.kg of feed-1.This experiment was carried out to evaluate the reproductive and zootechnical performance, and lipid deposition in the liver tissue of the Nile tilapia males fed diets containing different levels of digestible energy obtained from inclusion of soybean oil. Four hundred 400 broodfish (300 females and 100 males) were assigned to a completely randomized design composed of five levels of digestible energy (2,700, 2,950, 3,200, 3,450 and 3,700 kcal digestible energy.kg of feed-1) and four replications. The breeding were fed diets containing 35% of crude protein and submitted to hapas reproductive management for 101 days. The model adjusted by multiple regression analysis suggests the best result for sperm concentration and percentage of normal sperm for breeding fed diets containing 3.465.56 and 3.443.43 kcal digetible energy.kg of feed-1, producing 7.98 × 109 sperm.mL of semen-1 and 38.98% of normal sperm, respectively. The diet energy levels also caused an increase of lipid inclusion in the hepatocytes. The sperm production, seminal pH, sperm survival index and sperm activation time were not affected by energetic levels of the diets. The zootechnical performance was not affected by the energy levels of the diets either. The energy levels in the diets caused a linear increase in the lipid inclusion in the hepatocytes and affected the sperm quality by stimulating the spermatozoa production and the improving of normality indices of sperm morphology in energy levels around 3,450 kcal digestible energy.kg of feed-1.

Motilidade espermática, fertilização dos ovócitos e eclosão dos ovos de Jundiá em água contaminada por cádmio

The objective of this work was to evaluate the effect of water contaminated by cadmium on the rates of oocyte fertilization and egg hatching, and sperm motility duration on jundia catfish (Rhamdia quelen). Two trials were performed: in trial 1, it was evaluated the effect of cadmium on the artificial fertilization of oocytes and egg hatching and in trial 2, the effects of cadmium on the sperm motility duration were evaluated. In both trials, it was used a completely randomized experimental design with five levels of cadmium in the water (0; 20; 50; 80 and 110 mg L-1) evaluated with four (trial 1) or three replicates (trial 2). Fertilization and hatching rates showed linear response, remaining constant up to the levels of 28.6 and 40.4 mg L-1, respectively. From these levels, a linear effect inversely proportional between the fertilization and hatching rates and an increase in the levels of cadmium occurred. The increase on the level of cadium in the water had an inverse linear effect of sperm motility duration providing a reduction from 21.8s in clean water to 10.6s in contaminated water containing 110 mg L-1. Levels from 20.0 to 28.6 mg L-1 influence sperm and oocytes of jundia catfish, respectively.

Effects of the spermatozoa: oocyte ratio, water volume and water temperature on artificial fertilization and sperm activation of cascudo-preto

The objective of this study was to evaluate the effects of water volume and water temperature on the sperm motility duration and the number of spermatozoa, and the water volume on the fertilization rates of oocytes of Rhinelepis aspera. Experiments were carried out to evaluate the effect of semen dilutions (1.74×10-5, 1.74×10-4, 1.74×10-3, 1.74×10-2, 1.74×10-1 and 1.00 mL of sperm.mL-1 of water) and water temperature (5, 10, 15, 20, 25, 30, 35, 40, 45, and 50 oC) on spermatozoa motility duration. In addition, the effects of insemination dose (7×103, 7×104, 7×105, 7×106 and 7×107 spermatozoa.oocyte-1) and water volume (1.0, 30.0, 60.0, 90.0 and 120.0 mL water.2.0 mL-1 oocytes) on the artificial fertilization rates of oocytes were evaluated. The longest sperm motility duration were observed for the semen dilution of 1.74×10-5 mL semen.mL-1 water and in water at 5 oC. The highest fertilization rates were obtained for insemination doses between 7.00×103 and 1.23×107 spermatozoa. oocyte-1 and water volume of 28.11 mL water.2.0 mL-1 oocytes.

Inseminating dose and water volume applied to the artificial fertilization of Steindachneridion parahybae (Steindachner, 1877) (Siluriformes: Pimelodidae): Brazilian endangered fish

The Steindachneridion parahybae is an endangered catfish from Brazil and strategies applied for gametes optimization are necessary. The aim of this study was to assess inseminating doses and water volume upon the fertilization, hatching rates and percentage of normal larvae in S. parahybae . Was used a randomized design in factorial scheme (4×4) with four inseminating doses: 1.0×104, 1.0×105, 1.0×106, 1.0×107spermatozoa oocyte-1 and four volumes of water: 1, 35, 65 and 95mL of water g-1 of oocytes. The combination of doses and volumes were performed in triplicates (n=48). Each incubator (1.5L of useful volume) with 1g of oocytes was considered as an experimental unit. Significant interaction between inseminating doses and volumes of water to the values of the fertilization rates and quadratic effect of doses and volume for the values of hatching rates were observed. The doses and volumes did not influence the percentage of normal larvae (87.70±5.06%). It is recommended the use of 5.5×106 spermatozoa oocyte-1 and 1mL of water g-1 of oocytes during in vitro fertilization procedure. These results allowed us to develop new biotechnological strategies applied to the conservation of S. parahybae .

Colossoma macropomum females can reproduce more than once in the same reproductive period

The aim of the present study was to evaluate induced reproduction in Colossoma macropomum females at the beginning of the reproductive period and 75 days after the first spawning in which reproduction was induced. The experiment was conducted in Nova Mutum, MT, Brazil. Eight 4-year-old C. macropomum females with an average body weight of 6.7 ± 2.4 kg were used. Hormonal induction was performed at the beginning of the reproductive period and repeated 75 days after the first spawning. The following variables were then evaluated: weight of released oocytes, production index, absolute fecundity, oocyte diameter, fertilization rate, and hatching rate. Of the eight females that spawned during the first hormonal induction, three (37.5%) spawned again 75 days after the first spawning. Two females died after the first induced spawning. None of the means of the evaluated variables differed between the two induced spawnings, except for fertilization rate, which was greater (P <  0.05) with the first spawning (88.8 ± 6.1%) than in the second (74.1 ± 10.4%). The results of the present study indicate that C. macropomum females can reproduce again 75 days after a first induced spawning.

The requirement of digestible energy for females of Rhamdia quelen fed with ration based on vegetal ingredients

The present experiment was carried out to analyze the growth, the reproduction and the vigor of the offspring from females silver catfish ( Rhamdia quelen ) fed with ration pelleted containing different levels of digestible energy. The breeders were allocated in tanks under conditions of photoperiod and natural temperature. The fishes were fed for 255 days with isoproteic rations pelleted containing 35% of crude protein (CP) and rising levels of the digestible energy (DE) (2,700; 2,950; 3,200; 3,450; 3,700 kcal of DE.kg of ration -1 ). The fishes were distributed in a randomized experimental design compounded by five treatments and three repetitions. It was considered as one experimental unity a 16 m 2 tank containing six females and three males. The weight and the weight gain were evaluated. During the reproductive season the females were induced to breeding by hormonal manipulation and were evaluated the percentage of spawning females, the total fecundity, relative fecundity (number of oocytes per gram of spawning females), the fertilization ratio, the time to hatching and the vigor of larvae. The growth and reproductive parameters were not influenced (p>0.05) by the increasing levels of digestible energy of the rations. The feeding of females of Rhamdia quelen in breeding fit can be carried out with pelletized ration with 2,700 DE.kg of ration -1 based on vegetal ingredients, without damage to reproductive performance.