Adição da proteína específica do oviduto de porcas (pOSP) e da melatonina em meios de maturação e o efeito na clivagem in vitro de embriões suínos / Addition of the oviduct specific protein of porcine (pOSP) and melatonin in maturation media and their effect on the in vitro production of pig embryos

No presente estudo, utilizou-se a melatonina e a proteína específica do oviduto (pOSP) nos meios de maturação in vitro. Foram avaliadas a expansão do complexo cumulus-ovócito (CCOs), as concentrações intracelulares de espécies reativas de oxigênio (ROS) e o desenvolvimento embrionário nos diferentes grupos (C = controle; T1 = somente com melatonina; T2 = com melatonina e pOSP e T3 somente com pOSP). No tocante à expansão do CCOs, houve diferença (P<0,05) dos valores obtidos no grupo C em relação aos valores médios dos grupos T1, T2 e T3, porém não houve diferença entre os valores obtidos nos tratamentos (P>0,05). Na dosagem de ROS, não houve diferença entre os valores médios obtidos no grupo C (26,4±10,9) e o valor verificado no grupo T1 (23,4±7,8), porém no grupo T2 (21,3±9,7) o valor médio mostrou-se satisfatório em relação ao valor do grupo C. No entanto, o valor médio do grupo T3 (16,6±10,5) foi o que demonstrou resultado mais satisfatório quando comparado aos demais grupos (P<0,05). A produção de embriões foi avaliada por meio da taxa de clivagem. Não houve diferença (P >0,05) entre os valores obtidos entre o grupo C (48,9 %) e os valores verificados nos grupos T1 (51,5 %), T2 (50 %), T3 (57,7 %), nem destes entre si. Este estudo permitiu concluir que a proteína específica do oviduto recombinante e a melatonina foram eficientes em melhorar a expansão dos CCOs. Além disso, as células tratadas com pOSP mostraram-se com menor quantidade de ROS, podendo a pOSP ser considerada um antioxidante proteico.(AU)

The present study used melatonin and recombinant oviduct specific protein (pOSP) in in vitro maturation medium (IVM). The expansion of the cumulus-oocyte complexes (COCs), the intracellular concentrations of reactive oxygen species (ROS) and embryo development of the different groups were evaluated (C = control; T1 = melatonin; T2 = melatonin and pOSP and T3 = pOSP). Regarding the COCs expansion, the groups T1, T2 and T3 showed satisfactory results compared with group C (P<0.05), but there was no difference between treatments (P>0.05). In the ROS dosage, there was no difference between the mean values obtained in group C (26.4 ± 10.9) and group 1 (23.4 ± 7.8). However, in group 2 (21.3 ± 9.7), the average value was found to be satisfactory in relation group C. Despite that, the average value of treatment 3 (16.6 ± 10.5) was the most satisfactory result found compared to the other groups (P<0.05). The production of embryos was evaluated by cleavage rate, there was no difference between the values obtained in group C and the values recorded in groups T1 (51.5 %), T2 (50 %), T3 (57.7 %), and among them. This study showed that the pOSP and the melatonin were effective in the improvement of the expansion of COCs cells. In addition, the cells that were treated with pOSP presented a lower amount of ROS, allowing the pOSP to be considered a proteic antioxidant.(AU)

Adição da proteína específica do oviduto de porcas (pOSP) e da melatonina em meios de maturação e o efeito na clivagem in vitro de embriões suínos / Addition of the oviduct specific protein of porcine (pOSP) and melatonin in maturation media and their effect on the in vitro production of pig embryos

No presente estudo, utilizou-se a melatonina e a proteína específica do oviduto (pOSP) nos meios de maturação in vitro. Foram avaliadas a expansão do complexo cumulus-ovócito (CCOs), as concentrações intracelulares de espécies reativas de oxigênio (ROS) e o desenvolvimento embrionário nos diferentes grupos (C = controle; T1 = somente com melatonina; T2 = com melatonina e pOSP e T3 somente com pOSP). No tocante à expansão do CCOs, houve diferença (P<0,05) dos valores obtidos no grupo C em relação aos valores médios dos grupos T1, T2 e T3, porém não houve diferença entre os valores obtidos nos tratamentos (P>0,05). Na dosagem de ROS, não houve diferença entre os valores médios obtidos no grupo C (26,4±10,9) e o valor verificado no grupo T1 (23,4±7,8), porém no grupo T2 (21,3±9,7) o valor médio mostrou-se satisfatório em relação ao valor do grupo C. No entanto, o valor médio do grupo T3 (16,6±10,5) foi o que demonstrou resultado mais satisfatório quando comparado aos demais grupos (P<0,05). A produção de embriões foi avaliada por meio da taxa de clivagem. Não houve diferença (P >0,05) entre os valores obtidos entre o grupo C (48,9 %) e os valores verificados nos grupos T1 (51,5 %), T2 (50 %), T3 (57,7 %), nem destes entre si. Este estudo permitiu concluir que a proteína específica do oviduto recombinante e a melatonina foram eficientes em melhorar a expansão dos CCOs. Além disso, as células tratadas com pOSP mostraram-se com menor quantidade de ROS, podendo a pOSP ser considerada um antioxidante proteico.(AU)

The present study used melatonin and recombinant oviduct specific protein (pOSP) in in vitro maturation medium (IVM). The expansion of the cumulus-oocyte complexes (COCs), the intracellular concentrations of reactive oxygen species (ROS) and embryo development of the different groups were evaluated (C = control; T1 = melatonin; T2 = melatonin and pOSP and T3 = pOSP). Regarding the COCs expansion, the groups T1, T2 and T3 showed satisfactory results compared with group C (P<0.05), but there was no difference between treatments (P>0.05). In the ROS dosage, there was no difference between the mean values obtained in group C (26.4 ± 10.9) and group 1 (23.4 ± 7.8). However, in group 2 (21.3 ± 9.7), the average value was found to be satisfactory in relation group C. Despite that, the average value of treatment 3 (16.6 ± 10.5) was the most satisfactory result found compared to the other groups (P<0.05). The production of embryos was evaluated by cleavage rate, there was no difference between the values obtained in group C and the values recorded in groups T1 (51.5 %), T2 (50 %), T3 (57.7 %), and among them. This study showed that the pOSP and the melatonin were effective in the improvement of the expansion of COCs cells. In addition, the cells that were treated with pOSP presented a lower amount of ROS, allowing the pOSP to be considered a proteic antioxidant.(AU)

Assessment of different solutions and incubation times in the hypoosmotic swelling test of frozen/thawed semen in Piau boars

The objective of this study was to assess the effect of several solutions and incubation times in the hypoosmotic swelling test in predicting the in vitro quality of the frozen/thawed semen of Piau boars. Four samples of frozen semen from five different Piau boars were used. For the assessment of the hypoosmotic test, three incubation times were used (5, 30 and 60 min) in three different hypoosmotic solutions (BTS at 75 mOsm/Kg, sucrrose at 100 mOsm/Kg and fructose and sodium citrate at 100 mOsm/Kg), thus, samples were submitted to each hypoosmotic solution in three incubation times. The means of sperm motility spermatic vigor and live sperm by supravital staining for the assessed samples were respectively, 37.5 ± 7.2%, 2.8 ± 0.3 and 37.4 ± 7.8% in thawing and 15.5 ± 7.6%, 1.7 ± 0.5 and 22.6 ± 6.6% after 120 mm of thermoresistance test (P < 0.05). There was no difference between the mean values obtained for reactive sperm in the different incubation times and hypoosmotic solutions, and also no interaction between time ans solutions. The results obtained in the resent study demonstrated that quality of frozen/thawed semen, besides offering aliernatives for hypoosmotic solutions and different incubation times in performing the hypoosmotic test to predict the sperm plasmatic membrane quality in boats.

Assessment of different solutions and incubation times in the hypoosmotic swelling test of frozen/thawed semen in Piau boars

The objective of this study was to assess the effect of several solutions and incubation times in the hypoosmotic swelling test in predicting the in vitro quality of the frozen/thawed semen of Piau boars. Four samples of frozen semen from five different Piau boars were used. For the assessment of the hypoosmotic test, three incubation times were used (5, 30 and 60 min) in three different hypoosmotic solutions (BTS at 75 mOsm/Kg, sucrrose at 100 mOsm/Kg and fructose and sodium citrate at 100 mOsm/Kg), thus, samples were submitted to each hypoosmotic solution in three incubation times. The means of sperm motility spermatic vigor and live sperm by supravital staining for the assessed samples were respectively, 37.5 ± 7.2%, 2.8 ± 0.3 and 37.4 ± 7.8% in thawing and 15.5 ± 7.6%, 1.7 ± 0.5 and 22.6 ± 6.6% after 120 mm of thermoresistance test (P < 0.05). There was no difference between the mean values obtained for reactive sperm in the different incubation times and hypoosmotic solutions, and also no interaction between time ans solutions. The results obtained in the resent study demonstrated that quality of frozen/thawed semen, besides offering aliernatives for hypoosmotic solutions and different incubation times in performing the hypoosmotic test to predict the sperm plasmatic membrane quality in boats.(AU)

Teste de ligação de espermatozoides de suínos da raça Piau à membrana perivitelina da gema do ovo / Sperm-binding assay in Piau swine breeds to the hens egg perivitelline membrane

O objetivo deste trabalho foi avaliar a utilização do teste de ligação de espermatozoides à membrana perivitelina da gema do ovo de galinha como uma alternativa de predizer a qualidade espermática pós-descongelamento em suínos da raça Piau. Foram utilizadas cinco partidas de sêmen congelado de um varrão, as quais foram subdivididas em quatro tratamentos, de acordo com o meio (BTS ou B-TALP) e as concentrações espermáticas utilizadas (20 ou 40 x 106 espermatozoides/mL). Além disso, utilizaram-se 25 partidas de sêmen congelado de cinco varrões (cinco partidas de cada animal), sendo que 10 μL de sêmen na concentração de 20 x 106 espermatozoides/mL foram incubados em meio BTS ou B-TALP. No experimento 1, o número médio de espermatozoides aderidos/mm2 variou de 157,0 ± 81,3 a 249,4 ± 119,3 (P > 0,05). No experimento 2, o número médio de espermatozoides ligados/mm2 foi de 339,3 ± 255,5 e 275,4 ± 164,0, respectivamente, para os meios BTS e B-TALP, sem diferença entre eles (P > 0,05). Concluiu-se que a concentração espermática, assim como os meios de incubação utilizados, não interferiu na capacidade de adesão de espermatozoides à membrana perivitelina da gema do ovo em animais da raça Piau. (AU)

The objective of this study was to verify the use of sperm-binding assay in Piau swine breeds to the hens egg perivitelline membrane as an alternative to predict semen post-thawing fertilizing capability. It were used five matches of frozen semen from a boar, and the matches divided into four treatments, according to media (BTS or B-TALP) or sperm concentration (20 or 40 x 106 sperm/ml). Moreover, Twenty five frozen semen samples were used from five males (five semen samples from each male), using 10 μL of sperm with the concentration of 20 x 106 spermatozoa/mL and incubated BTS or B-TALP. In experiment 1, the average number of sperm-binding/mm2 varied from 157.0 ± 81.3 to 249.4 ± 119.3 (P > 0.05). In experiment two, The average number of sperm-binding/mm2 was 339.3 ± 255.5 and 275.4 ± 164.0, respectively for BTS and B-TALP media, with no difference between themselves (P > 0.05). It was concluded that sperm concentration, as well as the incubation media used did not affect the sperm adhesion to hens egg perivitelline membrane in Piau breed animals. (AU)

Teste de ligação de espermatozoides de suínos da raça Piau à membrana perivitelina da gema do ovo / Sperm-binding assay in Piau swine breeds to the hen’s egg perivitelline membrane

O objetivo deste trabalho foi avaliar a utilização do teste de ligação de espermatozoides à membrana perivitelina da gema do ovo de galinha como uma alternativa de predizer a qualidade espermática pós-descongelamento em suínos da raça Piau. Foram utilizadas cinco partidas de sêmen congelado de um varrão, as quais foram subdivididas em quatro tratamentos, de acordo com o meio (BTS ou B-TALP) e as concentrações espermáticas utilizadas (20 ou 40 x 106 espermatozoides/mL). Além disso, utilizaram-se 25 partidas de sêmen congelado de cinco varrões (cinco partidas de cada animal), sendo que 10 μL de sêmen na concentração de 20 x 106 espermatozoides/mL foram incubados em meio BTS ou B-TALP. No experimento 1, o número médio de espermatozoides aderidos/mm2 variou de 157,0 ± 81,3 a 249,4 ± 119,3 (P > 0,05). No experimento 2, o número médio de espermatozoides ligados/mm2 foi de 339,3 ± 255,5 e 275,4 ± 164,0, respectivamente, para os meios BTS e B-TALP, sem diferença entre eles (P > 0,05). Concluiu-se que a concentração espermática, assim como os meios de incubação utilizados, não interferiu na capacidade de adesão de espermatozoides à membrana perivitelina da gema do ovo em animais da raça Piau.

The objective of this study was to verify the use of sperm-binding assay in Piau swine breeds to the hen’s egg perivitelline membrane as an alternative to predict semen post-thawing fertilizing capability. It were used five matches of frozen semen from a boar, and the matches divided into four treatments, according to media (BTS or B-TALP) or sperm concentration (20 or 40 x 106 sperm/ml). Moreover, Twenty five frozen semen samples were used from five males (five semen samples from each male), using 10 μL of sperm with the concentration of 20 x 106 spermatozoa/mL and incubated BTS or B-TALP. In experiment 1, the average number of sperm-binding/mm2 varied from 157.0 ± 81.3 to 249.4 ± 119.3 (P > 0.05). In experiment two, The average number of sperm-binding/mm2 was 339.3 ± 255.5 and 275.4 ± 164.0, respectively for BTS and B-TALP media, with no difference between themselves (P > 0.05). It was concluded that sperm concentration, as well as the incubation media used did not affect the sperm adhesion to hen’s egg perivitelline membrane in Piau breed animals.