RESUMO.-Este estudo descreveu as características seminais, da membrana plasmática e do acrossoma de espermatozoide congelado/descongelado de 19 ejaculados de garanhões da raça Nordestina. Os aspectos analisados incluíram os parâmetros físicos do sêmen fresco; a motilidade e a longevidade do sêmen diluído e descongelado; a morfologia espermática, integridade funcional e estrutural da membrana plasmática do espermatozoide e a habilidade de ligação do espermatozoide à membrana perivitelina da gema do ovo de galinha do sêmen descongelado. This paper describes the seminal characteristics of the plasma membrane of frozenthawed sperm. Nineteen ejaculates of Nordestino horse breed. The aspects analyzed in the physical parameters of fresh semen were total and progressive motility and your longevity after dilution or thawed; sperm morphology, functional and structural integrity of the plasma membrane of the sperm and the sperm-binding ability to the perivitelline membrane of the yolk (MPV) after thawed. The variables were assessed by ANOVA with post hoc test of Student Newman-Keuls test (P<0.05). The total and progressive motility were higher in diluted semen than thawed (P<0.05). The average percentage of the major, minor and total defects was lower than the limit recommended by the CBRA. The percentage of reactive to hypo-osmotic swelling test was 14.21±1.12%, the intact membrane detected by supravitally test was 62.22±9.06% and the SYBR-14 was 81.47±26.9. The ability of sperm to bind to the MPV after thawing semen was 230.39±57.09. The total and progressive motility at time 0 min of termo resistance test was higher than to 150 minutes (P<0.05), and no difference was observed in the times 10 and 30 minutes. The results demonstrate that the use of additional laboratory tests help in the process of evaluation of samples, making possible to obtain more reliable and accurate information. Although cryopreservation has caused decrease in sperm motility and was used diluents with amides to diluted and cryopreservation protocol and this minimized the osmotic damage to sperm cells and maintained the morphological, functional and structural integrity of the plasma membrane of the sperm. These results are a reference for future studies since there are no comparative data on this breed.

ABSTRACT.-Souza W.L., Moraes E.A., Costa J.M.S., Souza P.H.F., Lopes Junior E.S., Oliveira R. The aim was to evaluate the effect of adding different concentrations of melatonin in ram semen diluted after cryopreservation. Ten ejaculates were collected0 from three adult ram (n=30) by means of artificial vagina for sheep. The collected samples were diluted in Tris-egg yolk, to a final concentration 200x10 6 sptz/mL kept in water bath at 32°C, and melatonin added as treatments: control; 100pM; 100nM; 100μM and 1mM melatonin. Then, the samples were cooled in a cold chamber at 5°C for two hours, in straws of 0.5mL and sealed. They were stored under the liquid nitrogen vapor for 15 minutes to 8cm of liquid blade and frozen with liquid nitrogen. Samples were analyzed for sperm motility, membrane integrity, acrosomal membrane, mitochondrial activity, oxidative stress and quantification of the binding capacity. The variables were subjected to analysis of variance and the means were compared by Tukey test at 5% probability. The total and progressive motility of thawed sperm were higher in samples treated with 100pM melatonin (62.99 and 45.07%, respectively; P<0.05) when compared to other treatments. The addition of different concentrations of melatonin in semen diluted with the exception of 1mM concentration, a higher percentage of cells with intact plasma membrane, as compared with the control (P<0.05). The percentage of sperm with acrosome membrane integrity was higher in the semen with 100pM melatonin (P<0.05) than the other treatments. The high mitochondrial activity was higher in spermatozoa treated with 100pM melatonin (69.30%; P<0.05). Addition of 100nM melatonin reduced the amount of TBARS after cryopreservation (2.84, P<0.05) when compared with the other treatments. After thawing, the number of sperm which bind to the perivitelline membrane was higher in the melatonin treated with 100pM (155,73; P<0.05). Therefore, melatonin addition the semen diluted can be useful to enhance the cryopreservation of sheep semen, improving fertilization rates through artificial insemination.

This study evaluated the effect of adding different concentration of cholesterol-loaded-cyclodextrin (CLC) on sperm quality after thawing. Thirty ejaculates were diluted, centrifuged and resuspended to 120 million cells/mL in a Tris diluent. Semen was treated with 0 (Control), 0.75, 1.5, 3.0, 4.5, 6.0 or 7.5 mg of CLC. Then, the samples were cooled at 4 °C for 2 h, diluted with Tris-egg yolk and 2% glycerol, packaged into 0.5 mL straws, frozen in liquid nitrogen (N 2 ) vapor for 20 min before being plunged into N 2 . Straws were thawed at 37 °C for 30 sec, and evaluated for thermal resistance test (TRT); progressive motility using CASA; hypoosmotic test and for binding capacity of sperm to perivitelline membrane (PM). The variables were determined using ANOVA at 5% probability. Higher percentage of motile sperm were maintained after thawing and TTR when 0.75 mg CLC was added, evaluated at 0, 60 and 120 min of incubation (50.4, 33.8 and 22.5%, respectively) compared to other treatments (P<0.05). The percentage of coiling was higher in sperm treated with 6.0 and 7.0 mg of CLC than other treatments (P<0.05). Addition of 0.75 mg CLCs also resulted in more sperm binding to the PM after cryopreservation than control sperm (166 vs 65; P<0.05). However, when the spermatozoa binding potential was determined on a motile sperm basis by dividing the average number of spermatozoa bound to PM for each bucks by the percentage of motile spermatozoa, CLC treatment provided higher binding efficiency (1.52) than control (1.00; P<0.05). In conclusion, CLCs improved the percentage of post-thaw of motility in caprine sperm as well as increased the number of sperm that bind to PM. Addition of 0.75 mg of CLC to caprine sperm prior to cryopreservation improved the quality sperm motility for up to 2 h.Keywords: cyclodextrin, frozen, membrane, spermatozoa, seminal quality. ResumoEste estudo teve como objetivo avaliar o efeito da adição do colesterol carreado com a ciclodextrina (CCC) sobre a melhoria da qualidade espermática após a descongelação. Trinta ejaculados foram diluídos, centrifugados e ressuspendidos com Tris para concentração de 120 x 10 6 células/mL. O sêmen foi tratado com 0, 0,75, 1,5, 3,0, 4,5, 6,0 ou 7,5 mg de CCC. Em seguida, as amostras foram resfriadas a 4°C durante 2 horas, diluídas com Tris-Gema de ovo e 2% de glicerol, envasadas e colocadas sobre o vapor do nitrogênio líquido (N 2 ) por 20 min e depois mergulhadas no N 2 . As amostras foram descongeladas a 37°C por 30s, e avaliadas quanto: o teste de termorresistência (TRT); motilidade progressiva utilizando CASA; teste hiposmótico e a capacidade de ligação dos espermatozoides à membrana perivitelina (MP). As variáveis foram analisadas por meio da ANOVA e os tratamentos comparados a 5% de probabilidade. A motilidade dos espermatozoides (50,4; 33,8 e 22,5%) foi maior nas amostras tratadas com 0,75 mg de CCC após 0, 60 e 120 min de incubação quando comparado com demais tratamentos (P<0,05). As amostras tratadas com 6,0 e 7,0 mg de CCC apresentaram maior dobram...

Aimed to evaluate the effect of adding antioxidants as ascorbic acid, melatonin and Trolox C to diluted semen of ram with oxidative stress to potenciate fertilization after cryopreservation. Ten samples collected were diluted in Tris-egg yolk to a final concentration of 200x10 6 sperm/mL and kept in a water bath at 32°C. Antioxidants were added as follows: 100µM melatonin (MEL) +.05% ascorbic acid (AA); 100µM of MEL + 90µL of Trolox C (TRO); 90µL of TRO + 0.05% AA; and 100µM of MEL0.05% AA + 90µL of TRO. Semen was cooled in a cold chamber at 5°C for two hours and packaged, sealed in 0.5mL straws, packaged under liquid nitrogen vapor (N 2 L), 8cm of water depth for 15 minutes, and then immersed in N 2 L. Samples were assayed for motility, integrity of the plasma membrane and acrosomal membrane, mitochondrial activity, binding assay and oxidative stress spermatozoa. The variables were analyzed by ANOVA and means compared by Tukey test (P<0.05). Percentage of total and progressive motility was higher for sperm treated with MEL+AA+TRO (67% and 49.89%), MEL+AA (64.37% and 45.61%) and MEL+TRO (61.65% and 41.15%) compared with the other treatments (P<0.05). The integrity of the plasma membrane and acrosome was higher for all semen treated with antioxidant associations compared with control (P<0.05). Mitochondrial activity was higher in sperm treated with MEL+AA+TRO compared all treatments (P<0.05). The number of sperm binding to perivitelline membrane was higher for semen treated with antioxidant associations compared with control; also sperm treated with MEL+AA+TRO demonstrated higher effect of all (P<0.05). No difference was observed between the treatments by oxidative stress sperm (P>0.05). The addition of melatonin, ascorbic acid and Trolox C in diluted semen of ram improves sperm quality after thawing.

Over the past decade, electricity consumption in Brazil grew faster than generation capacity. This situation obliged an urgent return to investment in the sector, and revitalization of the restructuring in the national electricity sector. In these circumstances, the use of renewable energy sources, such the biomass, became an option for decentralized electricity generation. Sugar cane bagasse is one of the most important biomasss residues for electricity generation. The present publication analyses an investment made in the expansion of the energy cogeneration system in an industry that produces sugar and alcohol, from sugar cane, considering the seasonal bagasse price, energy generation costs and a 10 year period. With the new cogeneration system the factory became self-sufficient in energy, with a saleable surplus of 21,240 MWh, at an average power of 4,000 kW. However, an economic analysis indicated that the best option would have been to maintain the original system and sell surplus bagasse at R$ 26.00/t.

RESUMO: Nesta pesquisa avaliou-se o efeito do colesterol sobre o sêmen de garanhões da raça Nordestina sobre a qualidade espermática. Vinte ejaculados de dois garanhões foram diluídos com BotuSemen e colesterol carreado pela ciclodextrina (CCC) adicionado no sêmen: controle, 0,75mg de CCC e 1,0mg de CCC/120x106 sptz/mL, e incubado a 26°C/15min. O sêmen foi diluído 1:5 (v/v) com diluente Lactose-gema de ovo e resfriado a 5°C/2h, envasado em palhetas de 0,5mL, e acondicionado sob vapor de nitrogênio líquido, e depois imersos. As amostras foram descongeladas (37 °C/30s) e avaliadas. As variáveis foram avaliadas com ANOVA e teste de Tukey (P<0,05). A motilidade total e progressiva foi maior (P<0,05) no sêmen tratado com CCC comparado as amostras do grupo controle, e CCC promoveu maior percentual (P<0,05) de motilidade total e progressiva durante as 3 horas de incubação. A percentagem de espermatozoides com viabilidade e integridade foi maior (P<0,05) no sêmen tratado com CCC (81,47 e 86,07%) comparado ao controle (72,12 e 70,19%). O número de espermatozoides reativos ao teste hiposmótico foi maior (P<0,05) nas amostras de sêmen tratadas com CCC comparado ao controle. Adição de colesterol no sêmen de garanhões Nordestino melhora a qualidade espermática apos a criopreservação.

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objetivou-se avaliar o efeito da dimetilformamida associada ou não ao glicerol no diluente de sêmen de caprinos antes da criopreservação sobre a longevidade e a funcionalidade da membrana espermática. Ejaculados de quatro caprinos foram coletados e diluídos com Tris diluente. Posteriormente, foram determinados os seguintes tratamentos experimentas: Controle; TG + 2% de dimetilformamida (DMF2); TG + 3% de DMF (DMF3); TG + 4% de DMF (DMF4); TG + 5% de DMF (DMF5); e TG + 2% de glicerol + 2% DMF (DMF2G). As amostras foram resfriadas a 5 °C/2 h, envasadas em palhetas de 0,5 mL e colocadas sob vapor de nitrogênio por 7 min, após este tempo as palhetas foram imersas no nitrogênio liquido. Os tratamentos foram descongelados a 37 ºC/30 segundos. As amostras foram analisadas quanto à longevidade da motilidade progressiva e quanto ao teste hiposmótico. As variáveis foram submetidas à análise de variância e as médias foram comparadas pelo teste de Tukey a 5% de probabilidade. A motilidade progressiva dos espermatozoides foi maior ao longo dos 120 min do TTR, nas amostras tratadas com DMF2, quando comparado às demais concentrações testadas (P<0,05). Observou-se que os espermatozoides de caprinos tratados com DMF2 antes da criopreservação, apresentaram maior percentual de células com conservação na funcionalidade da membrana plasmática quando comparado aos demais tratamentos (P<0,05). Em conclusão, o uso de 2% de dimetilformamida no diluente de sêmen caprino antes da criopreservação proporciona efeitos benéficos aos espermatozoides após descongelamento, sendo indicado como substituto do glicerol na criopreservação de espermatozoides de caprinos.