Vitamin E and reduced glutathione in Prochilodus lineatus (curimba) semen cryopreservation (Characiformes: Prochilodontidae)

This study investigated the addition of antioxidants vitamin E and reduced glutathione on curimba (Prochilodus lineatus) semen cryopreservation and compared sodium bicarbonate solution and distilled water as activators. The experiment was conducted at the environmental station of CEMIG, in Itutinga-MG, Brazil, between December/2009 and January/2010. Semen samples (n = 7) with semen motility above 80% were diluted in cryoprotectant solutions composed of 10% methanol, 15% lactose and containing different concentrations of antioxidants: 50 (VE50), 100 (VE100) and 250 (VE250) µM of vitamin E, and 0.5 (RG0.5), 1.0 (RG1.0) and 1.5 (RG1.5) mM of reduced glutathione. A solution without antioxidants was used as a control. The semen was diluted at a ratio of 1:4 (100 ìL semen:400 &#8733;L cryoprotectant solution). The toxicity of the solutions was evaluated by investigating semen motility after 10 min in the solution. The rest of the diluted semen was placed into 0.5 mL straws maintained in nitrogen vapour for 24 hours and packed into a nitrogen liquid cylinder for four days. The samples were thawed in a water bath at 60°C for 8 s and the rate (%) and duration (s) of semen activation with distilled water or sodium bicarbonate was evaluated. In the toxicity test, we found that vitamin E and reduced glutathione were not toxic to curimba semen at any of the tested concentrations (P>0.05). The duration of motility was longer (P<0.05) in semen activated with sodium bicarbonate 1% (163 ± 11 s), which was considered the best activator for semen under these conditions. No significant differences were found between the cryoprotectant solutions used after freezing (P>0.05). Thus, the antioxidants vitamin E and reduced glutathione did not improve the quality of cryopreserved curimba semen, but they did not cause toxic effects to the semen in natura and they did not decrease its quality during cryopreservation.<br>Este estudo avaliou a adição de antioxidantes vitamina E e glutationa reduzida no sêmen criopreservado de curimba (Prochilodus lineatus) e comparou solução de bicarbonato de sódio e água destilada como ativadores. O experimento foi conduzido na estação ambiental da CEMIG, em Itutinga-MG, entre Dezembro/2009 e Janeiro/2010. Sêmen de sete animais, com motilidade espermática acima de 80%, foi diluído em soluções crioprotetoras compostas por metanol 10% e lactose 15% em diferentes concentrações de antioxidantes: 50 (VE50), 100 (VE100) e 250 (VE250) µM de vitamina E, 0,5 (RG5.5), 1,0 (RG1.0) e 1,5 (RG1.5) mM glutationa reduzida e uma solução controle sem antioxidante. O sêmen foi diluído na proporção de 1:4 (100 µL de sêmen: 400 µL de solução crioprotetora). A toxicidade das soluções foi avaliada pela motilidade espermática após de 10 minutos em solução. O restante do sêmen diluído foi armazenado em palhetas de 0,5 mL mantidos em vapor de nitrogênio por 24 horas e estocado em cilindro de nitrogênio líquido por quatro dias. As amostras foram descongeladas em banho-maria a 60°C por 8 segundos e avaliada a taxa (%) e duração (s) pela ativação do sêmen com água destilada e bicarbonato de sódio a 1%. No teste de toxicidade, observamos que os antioxidantes da vitamina E e glutationa, nas diferentes concentrações, não foram tóxicos para o sêmen do curimba (P>0,05). A duração da motilidade foi maior (P<0,05) no sêmen ativado com bicarbonato de sódio, sendo o melhor ativador para o sêmen nestas condições. Não houve diferenças significativas entre as soluções crioprotetoras utilizadas para sêmen antes e após congelamento (P>0,05). Assim, os antioxidantes vitamina E e glutationa reduzida não melhoram a qualidade do sêmen criopreservado de curimba, mas não causam efeitos tóxicos para o sêmen in natura e criopreservados por não diminuir sua qualidade durante a criopreservação

Vitamin E and reduced glutathione in Prochilodus lineatus (curimba) semen cryopreservation (Characiformes: Prochilodontidae)

This study investigated the addition of antioxidants vitamin E and reduced glutathione on curimba (Prochilodus lineatus) semen cryopreservation and compared sodium bicarbonate solution and distilled water as activators. The experiment was conducted at the environmental station of CEMIG, in Itutinga-MG, Brazil, between December/2009 and January/2010. Semen samples (n = 7) with semen motility above 80% were diluted in cryoprotectant solutions composed of 10% methanol, 15% lactose and containing different concentrations of antioxidants: 50 (VE50), 100 (VE100) and 250 (VE250) µM of vitamin E, and 0.5 (RG0.5), 1.0 (RG1.0) and 1.5 (RG1.5) mM of reduced glutathione. A solution without antioxidants was used as a control. The semen was diluted at a ratio of 1:4 (100 ìL semen:400 ∝L cryoprotectant solution). The toxicity of the solutions was evaluated by investigating semen motility after 10 min in the solution. The rest of the diluted semen was placed into 0.5 mL straws maintained in nitrogen vapour for 24 hours and packed into a nitrogen liquid cylinder for four days. The samples were thawed in a water bath at 60°C for 8 s and the rate (%) and duration (s) of semen activation with distilled water or sodium bicarbonate was evaluated. In the toxicity test, we found that vitamin E and reduced glutathione were not toxic to curimba semen at any of the tested concentrations (P>0.05). The duration of motility was longer (P0.05). Thus, the antioxidants vitamin E and reduced glutathione did not improve the quality of cryopreserved curimba semen, but they did not cause toxic effects to the semen in natura and they did not decrease its quality during cryopreservation

Efeito da inclusão de diferentes tipos de óleo na dieta de varrões sobre a qualidade do sêmen "in natura" Effect of the inclusion of different types of oil in the diet of boars on the quality of semen "in natura"

O experimento foi realizado no Departamento de Zootecnia da Universidade Federal de Lavras-MG, com o objetivo de avaliar o desempenho reprodutivo de varrões alimentados com rações suplementadas com diferentes tipos de óleos como fontes de ácidos graxos essenciais. Foram utilizados 24 suínos híbridos (AGPIC 337), machos inteiros, com peso inicial médio de 152,7 kg, em um delineamento experimental em blocos ao acaso, com quatro tratamentos (T1 = controle; T2 = 3% óleo de linhaça; T3 = 3% óleo comercial PUFA® e T4 = 3% de óleo de peixe) e seis repetições, com um animal por unidade experimental. As rações foram isoenergéticas e isoprotéicas, formuladas à base de milho e farelo de soja. As variáveis analisadas foram parâmetros seminais como motilidade, vigor, volume, concentração, número total de células e anomalias espermáticas, avaliadas entre a quarta e a décima primeira semana do início do fornecimento das rações aos animais. Não houve diferenças significativas entre os tratamentos para motilidade, concentração e anormalidades espermáticas totais (P>0,05), exceto para alteração de cauda (P<0,05), cuja suplementação com óleo de peixe apresentou pior resultado. Por outro lado, o vigor espermático foi significativamente maior para o óleo de peixe (P<0,05). Para volume e número total de espermatozóides no ejaculado, melhores resultados foram obtidos com o óleo comercial PUFA® (P<0,05). Conclui-se que, nas avaliações do sêmen in natura, o uso de óleo comercial PUFA® e óleo de peixe nas rações de varrões demonstrou ser mais eficiente para a produção de células espermáticas e para a viabilidade destas células, respectivamente.<br>The experiment was conducted at Animal Science Department of University of Lavras-MG, with the objective to evaluate the quality of semem in natura of boars fed with rations supplemented with different sources of oils as sources of essential fat acids. Twenty four hybrid boars (AGPIC 337) were used, with initial average weight of 152,7 kg, assigned into a randomized complete blocks design, with four treatments (T1 = control; T2 = 3% linseed oil; T3 = 3% commercial oil PUFA® and T4 = 3% of fish oil) and six repetitions, being each animal as a experimental unit. The diets were isoenergetics and isoproteics, formulated with corn and soybean meal. The analyzed variables were seminal parameters as mobility, vigor, volume, concentration, total number of cells and anomalies spermatic, valued between four and eleventh week of the experimental period. The data showed any significant differences among the treatments for the motility, concentration and total anomalies spermatic (P>0,05), except for alteration of tail (P<0,05), whose supplementation with fish oil shown worse results. Also the spermatic vigor were significantly higher (P<0.05) for the boars fed rations with fish oil suplementation. The volume and total number of spermatozoid in the ejaculate, showed best results for those boars fed rations with commercial oil PUFA® (P<0,05). It was conclude that, the quality of the semen in natura, for boars fed rations he supplemented with commercial oil PUFA® and the fish oil showed to be more efficient for the parameters spermatic cells production and cells viability in the young boars

Toxicity of cryoprotectants on Prochilodus lineatus (Valenciennes, 1837) (curimba) embryos in an experimental incubator (Characiformes: Prochilodontidae)

This paper investigated the effect of cryoprotectant substances on Prochilodus lineatus embryos in an experimental incubator. The prospective study applied combinations of polyvinyl alcohol, hydroxyethyl cellulose, gelatin and fetal bovine serum with dimethyl sulfoxide and ethylene glycol in a new experimental incubator. The morphology of embryos, larval viability and the efficiency of experimental incubators in maintaining the quality of embryos were evaluated. This study demonstrates the efficient association between hydroxyethylcellulose and dimethyl sulfoxide as greater viability (p<0.05) was found for embryos (72.9 ± 23.9%). It should also be noted the permeation of cryoprotectants in embryos through the changes found in chorion diameter, embryo diameter and embryo volume comparing the treatments versus control group (water) (p<0.05), this results can help in future cryopreservation protocols. Although the temperature and oxygenation differed between the usual and experimental incubators (p<0.05), the results showed a high fertilization rate (79.6 ± 13.2%) for experimental incubators (p<0.05) which is sufficient for the maintenance of embryos in a cryoprotective environment and effectively allows experimentation for long periods with cryoprotectant substances. Cryopreservation of fish embryos has not been accomplished yet and new approaches are required for understanding the permeability of teleost embryos, especially in Brazilian native species.Este trabalho avaliou o efeito de substâncias crioprotetoras sobre embriões de Prochilodus lineatus em uma incubadora experimental. O estudo aplicou combinações de álcool polivinílico, hidroxietilcelulose, gelatina e soro fetal bovino com dimetilosulfóxido e etilenoglicol em uma nova incubadora experimental. Foram avaliadas a morfologia dos embriões, a viabilidade larval e a eficiência das incubadoras experimentais na manutenção da qualidade dos embriões. Este estudo demonstra a associação eficiente entre hidroxietilcelulose e dimetilsulfóxido pela maior viabilidade (p<0,05) encontrada para os embriões (72,9 ± 23,9%). Deve-se notar também a permeação dos crioprotetores nos embriões através das alterações encontradas no diâmetro córion, diâmetro do embrião e no volume do embrião comparando os tratamentos ao grupo controle (água) (p<0,05), estes resultados podem ajudar em futuros protocolos de criopreservação. Embora a temperatura e a oxigenação diferiram entre as incubadoras comuns e as experimentais (p<0,05), os resultados mostraram elevada taxa de fertilização (79,6 ± 13,2%) para incubadoras experimentais (p<0,05), o que é suficiente para a manutenção de embriões em ambiente crioprotetor e permite efetivamente a experimentação por longos períodos com substâncias crioprotetoras. A criopreservação de embriões de peixes ainda não foi realizada e novas abordagens são necessárias para a compreensão da permeabilidade dos embriões de teleósteos, especialmente em espécies nativas brasileiras.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Toxicity of cryoprotectants on Prochilodus lineatus (Valenciennes, 1837) (curimba) embryos in an experimental incubator (Characiformes: Prochilodontidae)

This paper investigated the effect of cryoprotectant substances on Prochilodus lineatus embryos in an experimental incubator. The prospective study applied combinations of polyvinyl alcohol, hydroxyethyl cellulose, gelatin and fetal bovine serum with dimethyl sulfoxide and ethylene glycol in a new experimental incubator. The morphology of embryos, larval viability and the efficiency of experimental incubators in maintaining the quality of embryos were evaluated. This study demonstrates the efficient association between hydroxyethylcellulose and dimethyl sulfoxide as greater viability (p<0.05) was found for embryos (72.9 ± 23.9%). It should also be noted the permeation of cryoprotectants in embryos through the changes found in chorion diameter, embryo diameter and embryo volume comparing the treatments versus control group (water) (p<0.05), this results can help in future cryopreservation protocols. Although the temperature and oxygenation differed between the usual and experimental incubators (p<0.05), the results showed a high fertilization rate (79.6 ± 13.2%) for experimental incubators (p<0.05) which is sufficient for the maintenance of embryos in a cryoprotective environment and effectively allows experimentation for long periods with cryoprotectant substances. Cryopreservation of fish embryos has not been accomplished yet and new approaches are required for understanding the permeability of teleost embryos, especially in Brazilian native species