Adição do complexo ciclodextrina-colesterol na criopreservação do sêmen caprino

Detalhes bibliográficos
Autor(a) principal: Santos, Madriano Christilis da Rocha
Data de Publicação: 2013
Tipo de documento: Tese
Idioma: por
Título da fonte: LOCUS Repositório Institucional da UFV
Texto Completo: http://locus.ufv.br/handle/123456789/1869
Resumo: The objective of this study was to analyze by spermatic motility and vigor the accuracy of six different of Ciclodextrin-Cholesterol Compound (CCC) concentrations on the spermatic motility and vigor effect after goat semen cryopreservation process and to evaluate the plasma membrane function and integrity of thawed goat semen. Chapter 2 The objective of this study was to evaluate the functionality and integrity of plasma membrane of thawed goat semen after addition of Ciclodextrin-Cholesterol Compound (CCC) diluted in tris glycerol or tris ethylene glycol + yolk hen s egg diluent to goat semen. And further, to evaluate the way of inclusion of the CCC (together with diluent or preincubated for 15 minutes). Chapter 1 Five Billy goats were used and five samples were collected from each and then fractionated in seven aliquots. One aliquot did not receive the CCC been considered a negative control (Tcontrol), the others received the CCC in the following concentrations: 0.5; 1.0; 1.5; 2.0; 2.5; 3.0 mg for spermatic concentration of [ ] = 200 * 106/mL, being called treatments: T0.5; T1.,0; T1.5; T2.0; T2.5; and T3.0, respectively. The spermatic motility and vigor and the supravital and hypoosmotic test were applied to the post-thaw semen. Chapter 2 Four billy goats were used and three samples were collected from each with a total of twelve ejaculates. Each ejaculate was evaluated and the ones good to be frozen by the minimum requirements of motility and vigor by the Brazilian College of Animal Reproduction (CBRA, 1998) were fractionated in six aliquots of 350 * 106 spermatozoa (enough for 7 doses with 50 * 106 spermatozoa/semen straw). Each aliquot received 1.0 mg of CCC in the following treatment: TGlycerol negative control for Tris glycerol diluent without CCC; TGlycerol + CCC Tris glycerol diluent + CCC; TGlycerol + CCC + preincubated CCC diluted in isosmotic solution (normal saline) 15 minutes before addittion of the Tris glycerol diluent; TEthyleno diluent without CCC;TEthylene glycol negative control for Tris ethyleno glycol glycol + CCC Tris ethylene glycol + yolk hen s egg diluent + CCC; TEthylene glycol + CCC + preincubated CCC diluted in isosmotic solution (normal saline) 15 minutes before addition of the Tris ethylene glycol + yolk hen s egg diluent. Chapter 1 Treatments were considered to be different if (P < 0.05). The spermatic motility was affected by all treatments (P<0.05), T1.0 e T0.5 values were higher (33.45% ± 1.07 and 32.59 ± 0.98, respectively) than 28.62 ± 1.05 for the Tcontrol and above the minimum value of 30%, required by CBRA (1998) for thawed semen. The motility for treatments: T1.5; T2.0; T2.5 and T3.0 were 27.41 ± 0.96; 26.55% ± 0.94; 25.89% ± 0.99 e 25.17% ± 1.06, respectively. The sperm vigor was not affected by the treatments(P>0.05), and their mean values were:Tcontrol(2.1 ± 0.07), T0.5(2.24 ±0.07), T1.0(2.4 ±0.08), T1.5(2.28 ±0.07), T2.0(2.13 ±0.07), T2.5(2.19 ±0.07), T3.0(2.26±0.09). The percentage of cells with functional membrane in the supravital test in T0.5 (41.5 ± 0.85) was higher (P<0.05) than Tcontrol (37.7 ± 0.94). The percentage of plasma membrane functionality did not change (P>0.05) among the treatments T2.0; T2.5; T1.0; T3.0 (38.06 ± 0.91; 38.06 ± 1.0; 36.33 ± 0.81; 35.54 ± 0.81; 34.77 ± 1.13, respectively) as compared to Tcontrol ones. The percentage of plasma membrane functionality for treatment T1.5 (30.65 ± 0.98) was lower (P<0.05) than all the others. No difference was found (P>0.05) in the integrity of plasma membrane by hypoosmotic test among Tcontrol (28.55 ± 1.01), T2.5 (24.47 ± 0.76), T0.5 (24.21 ± 0.82), T1.0 (23.74 ± 0.85), T1.5 (21.92 ± 1.18), T2.0 (21.24 ± 0.87), T3.0 (19.68 ± 0.88). Chapter 2 It was found difference among the averages of all treatments for the spermatic motility (P<0.05) TGlycerol preincubated (37.08 ± 2.17), TGlycerol TEthylene glycol + CCC + preincubated TEthylene glycol + CCC + CCC + (35.00 ± 1.51), TGlycerol (29.55 ± 2.81), (29.17 ± 4.52), TEthylene glycol + CCC (25.83 ± 3.98), (23.64 ± 4.10). The vigor of thawed semen was not affected by treatments used (P>0.05), whose averages are: TEthyleno glycol + CCC + preincubated (2.96± 0.31), TEthyleno glycoll + CCC (2.91 ± 0.12), TGlycerol + CCC + preincubated (2.75 ± 0.2), T Ethyleno glycol (2.59 ± 0.35), TGlycerol + CCC (2.29 ± 0.18), TGlycerol (2.22 ± 0.18). No difference was found (P>0.05) on the percentage plasma membrane integrity by hypoosmotic test on treatments TGlycerol CCC/preincubated (49.65 ± 5.9), TGlycerol + CCC/preincubated + CCC (52.0 ± 4), TEthylene glycol+ (45.05 ± 4.83), TGlycerol (44.8 ± 2.31), TEthylene glycol (44.39 ± 4.66), TEthylene glycol + CCC (39.6 ± 4.96). The functionality of the plasma membrane by supravital test was not affected by the treatments (P>0.05). The treatments averages were: TEthylene 3.82), TEthylene glycol + CCC (26.96 ± 4.73), TGlycerol glycol+ CCC/preincubated + CCC/preincubated (29.0 ± (24.5 ± 4.2), TGlycerol + CCC (23.54 ± 4.49), TGlycerol (20.79 ± 3.35), TEthylene glycol (19.45 ± 4.08). The ATP production by mitochondrial sheath was affected by treatmens (P<0.05) and theirs means were: TEthylene glycol + CCC (62.92 ± 8.34), TEthylene glycol+ CCC/preincubated (62.72 ± 8.79), TEthylene glycol (59.19 ± 9.57), TGlycerol + CCC/preincubated (11.92 ± 6.89), TGlycerol (11.64 ± 5.43), TGlycerol + CCC (7.76 ± 3.31). The acrosomal glycol (78.19 ± 5.82), glycol + CCC (70.68 ± 5.77), membrane integrity means by treatments were: TEthylene TEthylene glycol+ CCC/preincubated (74.43 ± 6.31), TEthylene TGlycerol (47.06 ± 4.31), TGlycerol + CCC (46.88 ± 6.58), TGlycerol + CCC/preincubated (44.6 ± 8.57) and were influenced by the treatments (P<0.05). The plasma membrane integrity by fluorescent probes (Propidium iodide + diethyl carboxyfluorescein) were not affected by treatments (P>0.05) and theirs means were: TGlycerol (63.98 ± 8.4), TGlycerol + CCC (63.08 ± 9.05), TGlycerol + CCC/preincubated (52.24 ± 9.47), T5 (52.09 ± 10.88), TEthylene glycol+ CCC/preincubated (44.37 ± 8.96), TEthylene glycol (37.19 ± 10.81).Chapter 1 It is concluded that the addition of 0.5 mg of CCC at spermatic concentration of 200 * 106/mL reduced the loss of integrity and functionality of the plasma membrane. However, the addition of 1.0 mg of CCC at the spermatic concentration 200 * 106/ mL showed better results for sperm cell motility in post-thawed semen. Chapter 2 It is concluded based on the results that the addittion of 1.0 mg of CCC on the spermatic concentration of 200 * 106/ mL did not affected the integrity neither the functionality of plasma membrane of goat sperm. The Tris + Ethyleno glycol + yolk egg hen s yolk diluent did not alter the functionality of plasma membrane or vigor.. Although it has a positive influence on the acrossomal membrane integrity, negatively affects the motility of the thawed sperm compared to solvent-based Tris- glycerol. The preincubation of sperm with CCC does not alter the characteristics of sperm vigor, and integrity and functionality of the plasma membrane, but positively affect the motility of the thawed sperm.
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spelling Santos, Madriano Christilis da Rochahttp://lattes.cnpq.br/8042592022583019Guimarães, José Domingoshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782270U6Carvalho, Giovanni Ribeiro dehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4723068Z6Torres, Ciro Alexandre Alveshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787213D4Veloso, Cristina Mattoshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4723663Z4Paula, Tarcízio Antônio Rego dehttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4701637D5Santos, Giancarlo Magalhães doshttp://lattes.cnpq.br/33650203924121702015-03-26T12:54:56Z2014-09-042015-03-26T12:54:56Z2013-08-21SANTOS, Madriano Christilis da Rocha. Addition of ciclodextrina-cholesterol compound on cryopreservation of goat semen. 2013. 96 f. Tese (Doutorado em Genética e Melhoramento de Animais Domésticos; Nutrição e Alimentação Animal; Pastagens e Forragicul) - Universidade Federal de Viçosa, Viçosa, 2013.http://locus.ufv.br/handle/123456789/1869The objective of this study was to analyze by spermatic motility and vigor the accuracy of six different of Ciclodextrin-Cholesterol Compound (CCC) concentrations on the spermatic motility and vigor effect after goat semen cryopreservation process and to evaluate the plasma membrane function and integrity of thawed goat semen. Chapter 2 The objective of this study was to evaluate the functionality and integrity of plasma membrane of thawed goat semen after addition of Ciclodextrin-Cholesterol Compound (CCC) diluted in tris glycerol or tris ethylene glycol + yolk hen s egg diluent to goat semen. And further, to evaluate the way of inclusion of the CCC (together with diluent or preincubated for 15 minutes). Chapter 1 Five Billy goats were used and five samples were collected from each and then fractionated in seven aliquots. One aliquot did not receive the CCC been considered a negative control (Tcontrol), the others received the CCC in the following concentrations: 0.5; 1.0; 1.5; 2.0; 2.5; 3.0 mg for spermatic concentration of [ ] = 200 * 106/mL, being called treatments: T0.5; T1.,0; T1.5; T2.0; T2.5; and T3.0, respectively. The spermatic motility and vigor and the supravital and hypoosmotic test were applied to the post-thaw semen. Chapter 2 Four billy goats were used and three samples were collected from each with a total of twelve ejaculates. Each ejaculate was evaluated and the ones good to be frozen by the minimum requirements of motility and vigor by the Brazilian College of Animal Reproduction (CBRA, 1998) were fractionated in six aliquots of 350 * 106 spermatozoa (enough for 7 doses with 50 * 106 spermatozoa/semen straw). Each aliquot received 1.0 mg of CCC in the following treatment: TGlycerol negative control for Tris glycerol diluent without CCC; TGlycerol + CCC Tris glycerol diluent + CCC; TGlycerol + CCC + preincubated CCC diluted in isosmotic solution (normal saline) 15 minutes before addittion of the Tris glycerol diluent; TEthyleno diluent without CCC;TEthylene glycol negative control for Tris ethyleno glycol glycol + CCC Tris ethylene glycol + yolk hen s egg diluent + CCC; TEthylene glycol + CCC + preincubated CCC diluted in isosmotic solution (normal saline) 15 minutes before addition of the Tris ethylene glycol + yolk hen s egg diluent. Chapter 1 Treatments were considered to be different if (P < 0.05). The spermatic motility was affected by all treatments (P<0.05), T1.0 e T0.5 values were higher (33.45% ± 1.07 and 32.59 ± 0.98, respectively) than 28.62 ± 1.05 for the Tcontrol and above the minimum value of 30%, required by CBRA (1998) for thawed semen. The motility for treatments: T1.5; T2.0; T2.5 and T3.0 were 27.41 ± 0.96; 26.55% ± 0.94; 25.89% ± 0.99 e 25.17% ± 1.06, respectively. The sperm vigor was not affected by the treatments(P>0.05), and their mean values were:Tcontrol(2.1 ± 0.07), T0.5(2.24 ±0.07), T1.0(2.4 ±0.08), T1.5(2.28 ±0.07), T2.0(2.13 ±0.07), T2.5(2.19 ±0.07), T3.0(2.26±0.09). The percentage of cells with functional membrane in the supravital test in T0.5 (41.5 ± 0.85) was higher (P<0.05) than Tcontrol (37.7 ± 0.94). The percentage of plasma membrane functionality did not change (P>0.05) among the treatments T2.0; T2.5; T1.0; T3.0 (38.06 ± 0.91; 38.06 ± 1.0; 36.33 ± 0.81; 35.54 ± 0.81; 34.77 ± 1.13, respectively) as compared to Tcontrol ones. The percentage of plasma membrane functionality for treatment T1.5 (30.65 ± 0.98) was lower (P<0.05) than all the others. No difference was found (P>0.05) in the integrity of plasma membrane by hypoosmotic test among Tcontrol (28.55 ± 1.01), T2.5 (24.47 ± 0.76), T0.5 (24.21 ± 0.82), T1.0 (23.74 ± 0.85), T1.5 (21.92 ± 1.18), T2.0 (21.24 ± 0.87), T3.0 (19.68 ± 0.88). Chapter 2 It was found difference among the averages of all treatments for the spermatic motility (P<0.05) TGlycerol preincubated (37.08 ± 2.17), TGlycerol TEthylene glycol + CCC + preincubated TEthylene glycol + CCC + CCC + (35.00 ± 1.51), TGlycerol (29.55 ± 2.81), (29.17 ± 4.52), TEthylene glycol + CCC (25.83 ± 3.98), (23.64 ± 4.10). The vigor of thawed semen was not affected by treatments used (P>0.05), whose averages are: TEthyleno glycol + CCC + preincubated (2.96± 0.31), TEthyleno glycoll + CCC (2.91 ± 0.12), TGlycerol + CCC + preincubated (2.75 ± 0.2), T Ethyleno glycol (2.59 ± 0.35), TGlycerol + CCC (2.29 ± 0.18), TGlycerol (2.22 ± 0.18). No difference was found (P>0.05) on the percentage plasma membrane integrity by hypoosmotic test on treatments TGlycerol CCC/preincubated (49.65 ± 5.9), TGlycerol + CCC/preincubated + CCC (52.0 ± 4), TEthylene glycol+ (45.05 ± 4.83), TGlycerol (44.8 ± 2.31), TEthylene glycol (44.39 ± 4.66), TEthylene glycol + CCC (39.6 ± 4.96). The functionality of the plasma membrane by supravital test was not affected by the treatments (P>0.05). The treatments averages were: TEthylene 3.82), TEthylene glycol + CCC (26.96 ± 4.73), TGlycerol glycol+ CCC/preincubated + CCC/preincubated (29.0 ± (24.5 ± 4.2), TGlycerol + CCC (23.54 ± 4.49), TGlycerol (20.79 ± 3.35), TEthylene glycol (19.45 ± 4.08). The ATP production by mitochondrial sheath was affected by treatmens (P<0.05) and theirs means were: TEthylene glycol + CCC (62.92 ± 8.34), TEthylene glycol+ CCC/preincubated (62.72 ± 8.79), TEthylene glycol (59.19 ± 9.57), TGlycerol + CCC/preincubated (11.92 ± 6.89), TGlycerol (11.64 ± 5.43), TGlycerol + CCC (7.76 ± 3.31). The acrosomal glycol (78.19 ± 5.82), glycol + CCC (70.68 ± 5.77), membrane integrity means by treatments were: TEthylene TEthylene glycol+ CCC/preincubated (74.43 ± 6.31), TEthylene TGlycerol (47.06 ± 4.31), TGlycerol + CCC (46.88 ± 6.58), TGlycerol + CCC/preincubated (44.6 ± 8.57) and were influenced by the treatments (P<0.05). The plasma membrane integrity by fluorescent probes (Propidium iodide + diethyl carboxyfluorescein) were not affected by treatments (P>0.05) and theirs means were: TGlycerol (63.98 ± 8.4), TGlycerol + CCC (63.08 ± 9.05), TGlycerol + CCC/preincubated (52.24 ± 9.47), T5 (52.09 ± 10.88), TEthylene glycol+ CCC/preincubated (44.37 ± 8.96), TEthylene glycol (37.19 ± 10.81).Chapter 1 It is concluded that the addition of 0.5 mg of CCC at spermatic concentration of 200 * 106/mL reduced the loss of integrity and functionality of the plasma membrane. However, the addition of 1.0 mg of CCC at the spermatic concentration 200 * 106/ mL showed better results for sperm cell motility in post-thawed semen. Chapter 2 It is concluded based on the results that the addittion of 1.0 mg of CCC on the spermatic concentration of 200 * 106/ mL did not affected the integrity neither the functionality of plasma membrane of goat sperm. The Tris + Ethyleno glycol + yolk egg hen s yolk diluent did not alter the functionality of plasma membrane or vigor.. Although it has a positive influence on the acrossomal membrane integrity, negatively affects the motility of the thawed sperm compared to solvent-based Tris- glycerol. The preincubation of sperm with CCC does not alter the characteristics of sperm vigor, and integrity and functionality of the plasma membrane, but positively affect the motility of the thawed sperm.Os objetivos do presente estudo foram: Capítulo 1 analisar, por meio da motilidade e do vigor espermáticos, a eficácia da adição de seis diferentes concentrações do complexo ciclodextrina-colesterol (CCC) em afetar a capacidade e a intensidade de movimentação das células espermáticas após o processo de congelamento do sêmen caprino, e avaliar a integridade e a funcionalidade da membrana plasmática do sêmen caprino descongelado. Capítulo 2 avaliar a integridade e a funcionalidade da membrana plasmática de espermatozoide após a adição do complexo ciclodextrina-colesterol (CCC) diluído em extensores à base de Tris-Glicerol ou à base de Tris-Etilenoglicol- gema de ovo ao sêmen caprino, e avaliar a forma de inclusão desse complexo (junto com o diluente ou pré-incubado por 15 minutos). No experimento descrito no Capítulo 1, cinco ejaculados foram coletados e fracionados de cinco reprodutores caprinos, saudáveis e com fertilidade comprovada. Uma fração foi utilizada como tratamento controle negativo (Tcontrole), onde não foi incluído o CCC. Adicionou-se nas demais alíquotas o diluente de congelamento (à base de Tris-glicerol) junto com o CCC nas concentrações de 0,5; 1,0; 1,5; 2,0; 2,5; e 3,0 mg, que foram consideradas como os tratamentos: T0,5; T1,0; T1,5; T2,0; T2,5; e T3,0, respectivamente, com uma concentração espermática de [ ] = 200 * 106/mL. Após o congelamento foram feitas as avaliações de motilidade e vigor espermáticos, o teste hiposmótico e o teste supravital. No experimento descrito no Capítulo 2, foram coletados três ejaculados de cada um dos quatro bodes, totalizando 12 ejaculados. Cada ejaculado foi avaliado, e aqueles considerados aptos para o congelamento, segundo os parâmetros mínimos de motilidade e vigor espermáticos exigidos pelo CBRA (1998), foram fracionados em seis alíquotas de 350 * 106 espermatozoides (quantidade suficiente para sete doses de 50 * 106 de espermatozoides/palheta de 0,25 mL). Cada alíquota recebeu 1,0 mg de CCC, nos seguintes tratamentos: TGlicerol controle negativo para o diluente à base de Tris-glicerol sem CCC; TGlicerol + CCC para o diluente à base de Tris-glicerol + CCC; TGlicerol + CCC/pré-incubado CCC diluído em solução isosmótica ao sêmen (soro fisiológico) com 15 minutos de incubação antes da adição do diluente à base de Tris-glicerol; TEtilenoglicol controle negativo para o diluente à base de Tris-Etilenoglicol-gema de ovo; TEtilenoglicol + CCC para o diluente à base de Tris-Etilenoglicol-gema de ovo + CCC, TEtilenoglicol+ incubado CCC/pré- para o CCC diluído em solução isoosmótica ao sêmen (soro fisiológico) com 15 minutos de incubação antes da adição do diluente à base de Tris- Etilenoglicol-gema de ovo. Capítulo 1 O valor de &#945; foi de 5% para todos os parâmetros avaliados. Constatou-se que todos os tratamentos apresentaram diferenças (P<0,05) para o parâmetro motilidade espermática e que os tratamentos T1,0 e T0,5 evidenciaram ser os maiores valores (33,45% ± 1,07 e 32,59% ± 0,98, respectivamente) em relação a 28,62% ± 1,05 do Tcontrole e estando acima do valor mínimo de 30%, preconizado pelo CBRA (1998) para sêmen descongelado. A motilidade espermática dos tratamentos T1,5; T2,5; T2,0; e T3,0 foram: 27,41% ± 0,96; 26,55% ± 0,94; 25,89% ± 0,99; e 25,17% ± 1,06, respectivamente. O vigor espermático não variou (P>0,05) entre tratamentos, sendo: T controle (2,1 ± 0,07), T0,5 (2,24 ± 0,07), T1,0 (2,4 ± 0,08), T1,5 (2,28 ± 0,07), T2,0 (2,13 ± 0,07), T2,5 (2,19 ± 0,07) e T3,0 (2,26 ± 0,09). O percentual de células com membrana funcional pelo teste supravital em T0,5 (41,5 ± 0,85) foi superior (P<0,05) ao do Tcontrole (37,7 ± 0,94), não tendo havido variações (P>0,05) nos percentuais médios entre os tratamentos: T2,0; T2,5; T1,0; e T3,0 (38,06 ± 0,91; 38,06 ± 1,0; 36,33 ± 0,81; 35,54 ± 0,81; e 34,77 ± 1,13, respectivamente), em relação ao do Tcontrole. O percentual de membrana funcional do tratamento T1,5 (30,65 ± 0,98) foi inferior (P<0,05) ao dos demais tratamentos. Não foram observadas diferenças (P>0,05) na integridade de membrana plasmática por meio do teste hiposmótico para Tcontrole (28,55 ± 1,01), T2,5 (24,47 ± 0,76), T0,5 (24,21 ± 0,82), T1,0 (23,74 ± 0,85), T1,5 (21,92 ± 1,18), T2,0 (21,24 ± 0,87) e T3,0 (19,68 ± 0,88), pelo teste de Kruskal-Wallis. Capítulo 2 O valor de &#945; foi de 5% para todos os parâmetros avaliados. As médias da motilidade espermática diferiram (P<0,05) entre todos os tratamentos TGlicerol + CCC/pré-incubado (37,08 ± 2,17), TGlicerol + CCC (35,00 ± 1,51), TGlicerol (29,55 ± 2,81), TEtilenoglicol+CCC/pré-incubado (29,17 ± 4,52), TEtilenoglicol + CCC (25,83 ± 3,98) e TEtilenoglicol (23,64 ± 4,10). O vigor espermático do sêmen descongelado não foi afetado pelos tratamentos aplicados (P>0,05), apresentando as seguintes médias TEtilenoglicol + CCC/pré-incubado (2,96 ± 0,31), TEtilenoglicol + CCC (2,91 ± 0,12), TGlicerol + CCC/pré-incubado (2,75 ± 0,2), TEtilenoglicol (2,59 ± 0,35), TGlicerol + CCC (2,29 ± 0,18), TGlicerol (2,22 ± 0,18). Não foram observadas diferenças (P>0,05) nos percentuais médios para integridade de membrana pelo teste hiposmótico dos tratamentos TGlicerol + CCC/pré-incubado (52,0 ± 4), TEtilenoglicol+ CCC/pré-incubado (49,65 ± 5,9), TGlicerol + CCC (45,05 ± 4,83), TGlicerol (44,8 ± 2,31), TEtilenoglicol (44,39 ± 4,66) e TEtilenoglicol + CCC (39,6 ± 4,96). Diferenças não foram observadas (P>0,05) entre as médias dos tratamentos TEtilenoglicol + CCC/pré- incubado (29,0 ± 3,82), TEtilenoglicol + CCC (26,96 ± 4,73), TGlicerol + CCC/pré-incubado (24,5 ± 4,2), TGlicerol + CCC (23,54 ± 4,49), TGlicerol (20,79 ± 3,35), TEtilenoglicol (19,45 ± 4,08), para funcionalidade da membrana plasmática, pelo teste supravital. Foram observadas diferenças (P<0,05) na produção de ATP na bainha mitocondrial nos tratamentos TEtilenoglicol + CCC (62,92 ± 8,34), TEtilenoglicol+ CCC/pré-incubado (62,72 ± 8,79), TEtilenoglicol (59,19 ± 9,57), TGlicerol (11,64 ± 5,43) e TGlicerol + CCC + CCC/pré-incubado (11,92 ± 6,89), TGlicerol (7,76 ± 3,31). Foram constatadas diferenças (P<0,05) na integridade de membrana acrossomal nos tratamentos TEtilenoglicol (78,19 ± 5,82), TEtilenoglicol+ CCC/pré-incubado (74,43 ± 6,31), TEtilenoglicol + CCC (70,68 ± 5,77), TGlicerol (47,06 ± 4,31), TGlicerol + CCC (46,88 ± 6,58) e TGlicerol + CCC/pré-incubado (44,6 ± 8,57). Não foram encontradas diferenças (P>0,05) na integridade de membrana plasmática por meio das sondas fluorescentes (IP Dietilcarboxifluorosceína) nos tratamentos TGlicerol (63,98 ± 8,4), TGlicerol (63,08 ± 9,05), TGlicerol + CCC/pré-incubado + + CCC (52,24 ± 9,47), T5 (52,09 ± 10,88), TEtilenoglicol+ CCC/pré-incubado (44,37 ± 8,96) e TEtilenoglicol (37,19 ± 10,81). Capítulo 1 Conclui-se que a adição de 0,5 mg do CCC ao sêmen com a concentração espermática de 200 * 106 /mL diminui as perdas de integridade e funcionalidade da membrana plasmática. No entanto, a adição de 1,0 mg de CCC ao sêmen com a concentração espermática de 200 * 106 /mL propicia melhor resultado na motilidade espermática após o descongelamento do sêmen. Capítulo 2 Diante dos resultados observados, conclui-se que a adição de 1,0 mg do complexo ciclodextrina-colesterol no sêmen com a concentração espermática de 200 * 106 /mL não altera a integridade e/ou a funcionalidade da membrana plasmática do espermatozoide caprino. O diluidor à base de Tris-gema de ovo- etileno glicol não altera a funcionalidade da membrana plasmática ou o vigor espermático,embora influencie positivamente a integridade da membrana acrossomal afete negativamente a motilidade espermática do sêmen descongelado, comparado ao diluente à base de Tris-Glicerol. A pré-incubação do sêmen com o complexo ciclodextrina-colesterol não altera as características de vigor espermático, nem a integridade e a funcionalidade da membrana plasmática dos espermatozoides, mas afeta positivamente a motilidade espermática do sêmen descongelado.Coordenação de Aperfeiçoamento de Pessoal de Nível Superiorapplication/pdfporUniversidade Federal de ViçosaDoutorado em ZootecniaUFVBRGenética e Melhoramento de Animais Domésticos; Nutrição e Alimentação Animal; Pastagens e ForragiculColesterolCaprinoSêmenCiclodextrinaCholesterolGoatsSemenCyclodextrinCNPQ::CIENCIAS AGRARIAS::ZOOTECNIAAdição do complexo ciclodextrina-colesterol na criopreservação do sêmen caprinoAddition of ciclodextrina-cholesterol compound on cryopreservation of goat semeninfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALtexto completo.pdfapplication/pdf540314https://locus.ufv.br//bitstream/123456789/1869/1/texto%20completo.pdf639cf7713b1947ed60a07182975bb9f8MD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain192332https://locus.ufv.br//bitstream/123456789/1869/2/texto%20completo.pdf.txt3a8c8a2eac12458c30d88be15e669089MD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3732https://locus.ufv.br//bitstream/123456789/1869/3/texto%20completo.pdf.jpg9919f3aea09e30bfd94ab31b9660a6a0MD53123456789/18692016-04-07 23:16:06.937oai:locus.ufv.br:123456789/1869Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-08T02:16:06LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.por.fl_str_mv Adição do complexo ciclodextrina-colesterol na criopreservação do sêmen caprino
dc.title.alternative.eng.fl_str_mv Addition of ciclodextrina-cholesterol compound on cryopreservation of goat semen
title Adição do complexo ciclodextrina-colesterol na criopreservação do sêmen caprino
spellingShingle Adição do complexo ciclodextrina-colesterol na criopreservação do sêmen caprino
Santos, Madriano Christilis da Rocha
Colesterol
Caprino
Sêmen
Ciclodextrina
Cholesterol
Goats
Semen
Cyclodextrin
CNPQ::CIENCIAS AGRARIAS::ZOOTECNIA
title_short Adição do complexo ciclodextrina-colesterol na criopreservação do sêmen caprino
title_full Adição do complexo ciclodextrina-colesterol na criopreservação do sêmen caprino
title_fullStr Adição do complexo ciclodextrina-colesterol na criopreservação do sêmen caprino
title_full_unstemmed Adição do complexo ciclodextrina-colesterol na criopreservação do sêmen caprino
title_sort Adição do complexo ciclodextrina-colesterol na criopreservação do sêmen caprino
author Santos, Madriano Christilis da Rocha
author_facet Santos, Madriano Christilis da Rocha
author_role author
dc.contributor.authorLattes.por.fl_str_mv http://lattes.cnpq.br/8042592022583019
dc.contributor.author.fl_str_mv Santos, Madriano Christilis da Rocha
dc.contributor.advisor-co1.fl_str_mv Guimarães, José Domingos
dc.contributor.advisor-co1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4782270U6
dc.contributor.advisor-co2.fl_str_mv Carvalho, Giovanni Ribeiro de
dc.contributor.advisor-co2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4723068Z6
dc.contributor.advisor1.fl_str_mv Torres, Ciro Alexandre Alves
dc.contributor.advisor1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4787213D4
dc.contributor.referee1.fl_str_mv Veloso, Cristina Mattos
dc.contributor.referee1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4723663Z4
dc.contributor.referee2.fl_str_mv Paula, Tarcízio Antônio Rego de
dc.contributor.referee2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4701637D5
dc.contributor.referee3.fl_str_mv Santos, Giancarlo Magalhães dos
dc.contributor.referee3Lattes.fl_str_mv http://lattes.cnpq.br/3365020392412170
contributor_str_mv Guimarães, José Domingos
Carvalho, Giovanni Ribeiro de
Torres, Ciro Alexandre Alves
Veloso, Cristina Mattos
Paula, Tarcízio Antônio Rego de
Santos, Giancarlo Magalhães dos
dc.subject.por.fl_str_mv Colesterol
Caprino
Sêmen
Ciclodextrina
topic Colesterol
Caprino
Sêmen
Ciclodextrina
Cholesterol
Goats
Semen
Cyclodextrin
CNPQ::CIENCIAS AGRARIAS::ZOOTECNIA
dc.subject.eng.fl_str_mv Cholesterol
Goats
Semen
Cyclodextrin
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS AGRARIAS::ZOOTECNIA
description The objective of this study was to analyze by spermatic motility and vigor the accuracy of six different of Ciclodextrin-Cholesterol Compound (CCC) concentrations on the spermatic motility and vigor effect after goat semen cryopreservation process and to evaluate the plasma membrane function and integrity of thawed goat semen. Chapter 2 The objective of this study was to evaluate the functionality and integrity of plasma membrane of thawed goat semen after addition of Ciclodextrin-Cholesterol Compound (CCC) diluted in tris glycerol or tris ethylene glycol + yolk hen s egg diluent to goat semen. And further, to evaluate the way of inclusion of the CCC (together with diluent or preincubated for 15 minutes). Chapter 1 Five Billy goats were used and five samples were collected from each and then fractionated in seven aliquots. One aliquot did not receive the CCC been considered a negative control (Tcontrol), the others received the CCC in the following concentrations: 0.5; 1.0; 1.5; 2.0; 2.5; 3.0 mg for spermatic concentration of [ ] = 200 * 106/mL, being called treatments: T0.5; T1.,0; T1.5; T2.0; T2.5; and T3.0, respectively. The spermatic motility and vigor and the supravital and hypoosmotic test were applied to the post-thaw semen. Chapter 2 Four billy goats were used and three samples were collected from each with a total of twelve ejaculates. Each ejaculate was evaluated and the ones good to be frozen by the minimum requirements of motility and vigor by the Brazilian College of Animal Reproduction (CBRA, 1998) were fractionated in six aliquots of 350 * 106 spermatozoa (enough for 7 doses with 50 * 106 spermatozoa/semen straw). Each aliquot received 1.0 mg of CCC in the following treatment: TGlycerol negative control for Tris glycerol diluent without CCC; TGlycerol + CCC Tris glycerol diluent + CCC; TGlycerol + CCC + preincubated CCC diluted in isosmotic solution (normal saline) 15 minutes before addittion of the Tris glycerol diluent; TEthyleno diluent without CCC;TEthylene glycol negative control for Tris ethyleno glycol glycol + CCC Tris ethylene glycol + yolk hen s egg diluent + CCC; TEthylene glycol + CCC + preincubated CCC diluted in isosmotic solution (normal saline) 15 minutes before addition of the Tris ethylene glycol + yolk hen s egg diluent. Chapter 1 Treatments were considered to be different if (P < 0.05). The spermatic motility was affected by all treatments (P<0.05), T1.0 e T0.5 values were higher (33.45% ± 1.07 and 32.59 ± 0.98, respectively) than 28.62 ± 1.05 for the Tcontrol and above the minimum value of 30%, required by CBRA (1998) for thawed semen. The motility for treatments: T1.5; T2.0; T2.5 and T3.0 were 27.41 ± 0.96; 26.55% ± 0.94; 25.89% ± 0.99 e 25.17% ± 1.06, respectively. The sperm vigor was not affected by the treatments(P>0.05), and their mean values were:Tcontrol(2.1 ± 0.07), T0.5(2.24 ±0.07), T1.0(2.4 ±0.08), T1.5(2.28 ±0.07), T2.0(2.13 ±0.07), T2.5(2.19 ±0.07), T3.0(2.26±0.09). The percentage of cells with functional membrane in the supravital test in T0.5 (41.5 ± 0.85) was higher (P<0.05) than Tcontrol (37.7 ± 0.94). The percentage of plasma membrane functionality did not change (P>0.05) among the treatments T2.0; T2.5; T1.0; T3.0 (38.06 ± 0.91; 38.06 ± 1.0; 36.33 ± 0.81; 35.54 ± 0.81; 34.77 ± 1.13, respectively) as compared to Tcontrol ones. The percentage of plasma membrane functionality for treatment T1.5 (30.65 ± 0.98) was lower (P<0.05) than all the others. No difference was found (P>0.05) in the integrity of plasma membrane by hypoosmotic test among Tcontrol (28.55 ± 1.01), T2.5 (24.47 ± 0.76), T0.5 (24.21 ± 0.82), T1.0 (23.74 ± 0.85), T1.5 (21.92 ± 1.18), T2.0 (21.24 ± 0.87), T3.0 (19.68 ± 0.88). Chapter 2 It was found difference among the averages of all treatments for the spermatic motility (P<0.05) TGlycerol preincubated (37.08 ± 2.17), TGlycerol TEthylene glycol + CCC + preincubated TEthylene glycol + CCC + CCC + (35.00 ± 1.51), TGlycerol (29.55 ± 2.81), (29.17 ± 4.52), TEthylene glycol + CCC (25.83 ± 3.98), (23.64 ± 4.10). The vigor of thawed semen was not affected by treatments used (P>0.05), whose averages are: TEthyleno glycol + CCC + preincubated (2.96± 0.31), TEthyleno glycoll + CCC (2.91 ± 0.12), TGlycerol + CCC + preincubated (2.75 ± 0.2), T Ethyleno glycol (2.59 ± 0.35), TGlycerol + CCC (2.29 ± 0.18), TGlycerol (2.22 ± 0.18). No difference was found (P>0.05) on the percentage plasma membrane integrity by hypoosmotic test on treatments TGlycerol CCC/preincubated (49.65 ± 5.9), TGlycerol + CCC/preincubated + CCC (52.0 ± 4), TEthylene glycol+ (45.05 ± 4.83), TGlycerol (44.8 ± 2.31), TEthylene glycol (44.39 ± 4.66), TEthylene glycol + CCC (39.6 ± 4.96). The functionality of the plasma membrane by supravital test was not affected by the treatments (P>0.05). The treatments averages were: TEthylene 3.82), TEthylene glycol + CCC (26.96 ± 4.73), TGlycerol glycol+ CCC/preincubated + CCC/preincubated (29.0 ± (24.5 ± 4.2), TGlycerol + CCC (23.54 ± 4.49), TGlycerol (20.79 ± 3.35), TEthylene glycol (19.45 ± 4.08). The ATP production by mitochondrial sheath was affected by treatmens (P<0.05) and theirs means were: TEthylene glycol + CCC (62.92 ± 8.34), TEthylene glycol+ CCC/preincubated (62.72 ± 8.79), TEthylene glycol (59.19 ± 9.57), TGlycerol + CCC/preincubated (11.92 ± 6.89), TGlycerol (11.64 ± 5.43), TGlycerol + CCC (7.76 ± 3.31). The acrosomal glycol (78.19 ± 5.82), glycol + CCC (70.68 ± 5.77), membrane integrity means by treatments were: TEthylene TEthylene glycol+ CCC/preincubated (74.43 ± 6.31), TEthylene TGlycerol (47.06 ± 4.31), TGlycerol + CCC (46.88 ± 6.58), TGlycerol + CCC/preincubated (44.6 ± 8.57) and were influenced by the treatments (P<0.05). The plasma membrane integrity by fluorescent probes (Propidium iodide + diethyl carboxyfluorescein) were not affected by treatments (P>0.05) and theirs means were: TGlycerol (63.98 ± 8.4), TGlycerol + CCC (63.08 ± 9.05), TGlycerol + CCC/preincubated (52.24 ± 9.47), T5 (52.09 ± 10.88), TEthylene glycol+ CCC/preincubated (44.37 ± 8.96), TEthylene glycol (37.19 ± 10.81).Chapter 1 It is concluded that the addition of 0.5 mg of CCC at spermatic concentration of 200 * 106/mL reduced the loss of integrity and functionality of the plasma membrane. However, the addition of 1.0 mg of CCC at the spermatic concentration 200 * 106/ mL showed better results for sperm cell motility in post-thawed semen. Chapter 2 It is concluded based on the results that the addittion of 1.0 mg of CCC on the spermatic concentration of 200 * 106/ mL did not affected the integrity neither the functionality of plasma membrane of goat sperm. The Tris + Ethyleno glycol + yolk egg hen s yolk diluent did not alter the functionality of plasma membrane or vigor.. Although it has a positive influence on the acrossomal membrane integrity, negatively affects the motility of the thawed sperm compared to solvent-based Tris- glycerol. The preincubation of sperm with CCC does not alter the characteristics of sperm vigor, and integrity and functionality of the plasma membrane, but positively affect the motility of the thawed sperm.
publishDate 2013
dc.date.issued.fl_str_mv 2013-08-21
dc.date.available.fl_str_mv 2014-09-04
2015-03-26T12:54:56Z
dc.date.accessioned.fl_str_mv 2015-03-26T12:54:56Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/doctoralThesis
format doctoralThesis
status_str publishedVersion
dc.identifier.citation.fl_str_mv SANTOS, Madriano Christilis da Rocha. Addition of ciclodextrina-cholesterol compound on cryopreservation of goat semen. 2013. 96 f. Tese (Doutorado em Genética e Melhoramento de Animais Domésticos; Nutrição e Alimentação Animal; Pastagens e Forragicul) - Universidade Federal de Viçosa, Viçosa, 2013.
dc.identifier.uri.fl_str_mv http://locus.ufv.br/handle/123456789/1869
identifier_str_mv SANTOS, Madriano Christilis da Rocha. Addition of ciclodextrina-cholesterol compound on cryopreservation of goat semen. 2013. 96 f. Tese (Doutorado em Genética e Melhoramento de Animais Domésticos; Nutrição e Alimentação Animal; Pastagens e Forragicul) - Universidade Federal de Viçosa, Viçosa, 2013.
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dc.publisher.none.fl_str_mv Universidade Federal de Viçosa
dc.publisher.program.fl_str_mv Doutorado em Zootecnia
dc.publisher.initials.fl_str_mv UFV
dc.publisher.country.fl_str_mv BR
dc.publisher.department.fl_str_mv Genética e Melhoramento de Animais Domésticos; Nutrição e Alimentação Animal; Pastagens e Forragicul
publisher.none.fl_str_mv Universidade Federal de Viçosa
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reponame_str LOCUS Repositório Institucional da UFV
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