Effect of fortification of extender with α-terpineol and rosemary essential oil on post-thaw ovine semsen quality.

The goal of the present study was to assess the viability of the cryopreserved ovine (Ovis aries) semen, upon supplementation with α-terpineol and rosemary essential oil (Rosmarinus officinalis). The collection of the semen from six rams formed the pool, collected once a week during 7 weeks. The diluted semen was packed into straws (0.25 ml) and frozen in a TK 3000® device. Both α-terpineol and rosemary essential oil were added in the concentrations of 6.25, 12.5 and 25 µg/ml to the TRIS-yolk extender forming six experimental groups; the control group received only the TRIS-yolk extender. The samples were analyzed after thawing regarding motility and vigor, integrity of the plasmatic membrane, thermoresistance test (TT), mitochondrial membrane potential, acrosomal integrity and computer-assisted semen analysis (CASA). The levels of the thiobarbituric acid reactive substances were also analyzed. According to the results obtained with the addition of the concentrations of 6.25, 12.5 and 25 µg/ml of α-terpineol, significantly reduced the parameters assessed through CASA (VSL, LIN and WOB) and TT. Rosemary essential oil did not have deleterious effects on the spermatozoa and did not reduce the oxidative stress in the concentrations studied, although it presented absolute values higher than those of the control in several parameters. Alpha-terpineol in the concentrations studied was not able to reduce the oxidative stress and had toxic effect over the ovine spermatozoa.

Suplementação do ácido oleico no dilente seminal sobre o potencial mitocondrial de espermatozoides caprinos criopreservados / Supplementation of oleic acid to the seminal diluter on the mitochondrial potential of cryopreserved caprine sperm

The research aimed to evaluate the antioxidant effect of supplementation of 0.5μM, 5μM and 50μM of oleic acid to the TRIS-yolk extender on the mitochondrial potential (MIT) during the cryopreservation of goat sperm. For that, four Anglo-nubian goats were used, in which five samples / animal were collected, using artificial vagina. After evaluating the swirling and motility of the ejaculates, the pool was made, then diluted in TRIS-Gem and divided according to the treatments. After processing, the samples were packaged in 0.25mL straws and cryopreserved using the TK 3000® machine. After a minimum of 5 days of storage in a cryogenic cylinder, thawing was performed to assess the MIT of goat sperm after cryopreservation, using the lipophilic cationic fluorochrome JC-1. The data were submitted to analysis of variance (ANOVA), using the general linear models procedure (Proc GLM), and the Duncan test was used to compare the averages, with a 5% probability. The analyzes were performed using the Statistical Analysis System program (SAS Institute Inc, 2013). Thus, it was observed that the concentrations of 0.5μM and 5μM of oleic acid maintained the mitochondrial potential similar to the control, differing (p<0.05) only the concentration of 50μM. It can be concluded that 0.5μM and 5μM oleic acid are able to maintain the mitochondrial potential, prolonging the viability of cryopreserved goat sperm.

Influência de um ácido graxo adicionado ao diluente seminal sobre a integridade da membrana plasmática de espermatozoides caprinos criopreservados / Influence of a fatty acid added to the seminal extender on the integrity of the plasmatic membrane of cryopreserved caprine sperm

The objective of this study was to evaluate the antioxidant effects of supplementing different concentrations (0.5μM, 5μM and 50μM) of polyunsaturated fatty acid, arachidonic acid to the TRIS-yolk diluter on the integrity of the plasma membrane during the cryopreservation of goat sperm. For this purpose, four Anglo-nubian goats were used, in which five samples / animal were collected, using artificial vagina. After evaluating the swirling and motility of the ejaculates, the pool was made, then diluted in TRIS-Gem and divided according to the treatments. After processing, the samples were packaged in 0.25mL straws and cryopreserved using the TK3000® machine. Defrosting occurred after at least 5 days of storage in a cryogenic cylinder. Then, the integrity of the plasma membrane of goat sperm post cryopreservation was carried out, using the double staining method, where carboxyfluorescein diacetate (DCF) and propidium iodide (IP) were used. The data were analyzed and the results of the researched variable were subjected to analysis of variance (ANOVA) using the general linear models procedure (Proc GLM) and the Duncan test was used to compare the means, with a 5% probability. The analyzes were performed using the Statistical Analysis System program (SAS Institute Inc, 2013). After analysis, it was observed that the control group had the best percentage, and differed significantly (p<0.05) from the treatment with 50μM of arachidonic acid. It was concluded that the 50μM arachidonic acid concentration is not effective to maintain the integrity of the plasma membrane, and to minimize the oxidative stress of cryopreservation.

Influência de um ácido graxo adicionado ao diluente seminal sobre a integridade da membrana plasmática de espermatozoides caprinos criopreservados / Influence of a fatty acid added to the seminal extender on the integrity of the plasmatic membrane of cryopreserved caprine sperm

The objective of this study was to evaluate the antioxidant effects of supplementing different concentrations (0.5μM, 5μM and 50μM) of polyunsaturated fatty acid, arachidonic acid to the TRIS-yolk diluter on the integrity of the plasma membrane during the cryopreservation of goat sperm. For this purpose, four Anglo-nubian goats were used, in which five samples / animal were collected, using artificial vagina. After evaluating the swirling and motility of the ejaculates, the pool was made, then diluted in TRIS-Gem and divided according to the treatments. After processing, the samples were packaged in 0.25mL straws and cryopreserved using the TK3000® machine. Defrosting occurred after at least 5 days of storage in a cryogenic cylinder. Then, the integrity of the plasma membrane of goat sperm post cryopreservation was carried out, using the double staining method, where carboxyfluorescein diacetate (DCF) and propidium iodide (IP) were used. The data were analyzed and the results of the researched variable were subjected to analysis of variance (ANOVA) using the general linear models procedure (Proc GLM) and the Duncan test was used to compare the means, with a 5% probability. The analyzes were performed using the Statistical Analysis System program (SAS Institute Inc, 2013). After analysis, it was observed that the control group had the best percentage, and differed significantly (p<0.05) from the treatment with 50μM of arachidonic acid. It was concluded that the 50μM arachidonic acid concentration is not effective to maintain the integrity of the plasma membrane, and to minimize the oxidative stress of cryopreservation.(AU)

Suplementação do ácido oleico no dilente seminal sobre o potencial mitocondrial de espermatozoides caprinos criopreservados / Supplementation of oleic acid to the seminal diluter on the mitochondrial potential of cryopreserved caprine sperm

The research aimed to evaluate the antioxidant effect of supplementation of 0.5μM, 5μM and 50μM of oleic acid to the TRIS-yolk extender on the mitochondrial potential (MIT) during the cryopreservation of goat sperm. For that, four Anglo-nubian goats were used, in which five samples / animal were collected, using artificial vagina. After evaluating the swirling and motility of the ejaculates, the pool was made, then diluted in TRIS-Gem and divided according to the treatments. After processing, the samples were packaged in 0.25mL straws and cryopreserved using the TK 3000® machine. After a minimum of 5 days of storage in a cryogenic cylinder, thawing was performed to assess the MIT of goat sperm after cryopreservation, using the lipophilic cationic fluorochrome JC-1. The data were submitted to analysis of variance (ANOVA), using the general linear models procedure (Proc GLM), and the Duncan test was used to compare the averages, with a 5% probability. The analyzes were performed using the Statistical Analysis System program (SAS Institute Inc, 2013). Thus, it was observed that the concentrations of 0.5μM and 5μM of oleic acid maintained the mitochondrial potential similar to the control, differing (p<0.05) only the concentration of 50μM. It can be concluded that 0.5μM and 5μM oleic acid are able to maintain the mitochondrial potential, prolonging the viability of cryopreserved goat sperm.(AU)

Influência do tipo de parto na determinação sérica de glicose em cordeiros / Delivery type of influence on serum glucose determination in lambs

This study aimed to evaluate the effect of turn and type of birth on the behavior of the concentration ofglucose in Dorper lambs grazing in Piauí semiarid. 18 lambs were used in the Dorper, born aged twenty fourhours. For the evaluation of plasma concentrations of glucose, the animals were randomly distributed in shifts(day and night) and the type of birth (single and double). Blood samples were collected at 24 hours of elapsedtime after birth, soon after placed in tubes with sodium fluoride and taken to the Clinical Pathology Laboratoryof the University Veterinary Hospital - HVU, UFPI. Analysis of variance revealed a significant effect of theinteraction shift in the types of twin births (116.64 ± 6.28) on the concentration of plasma glucose. Theconclusion of this research was that the performance of twin lambs, is due to the amount of ingested colostrumdoes not depend directly on the amount of offspring or due to their own birth order.(AU)

Efeito da somatotropina bovina recombinante (rbST) sobre a concentração espermática de ovinos Santa Inês / Effect of recombinant bovine somatotropin (rbST) in sperm concentration of sheep Santa Inês

This study aimed to evaluate among the seminal characteristics, sperm concentration of Santa Inessheep, aged 22-36 months treated with recombinant bovine somatotropin (rbST). Divided into groups randomly:GI (2mL / 0.9% NaCl), G-II (100mg / rbST) and G-III (125mg / rbST), receiving treatments subcutaneouslyevery 14 days (D0,14,28, 42,56,70). They were measured scrototesticular biometrics. The ejaculates werecollected by artificial vagina, with dummy sheep, and analyzed for volume, vortex, motility, vigor, morphologyand sperm concentration. There was no significant difference (P > 0.05) in PE (30,1cm) and in almost all semenparameters. However, on sperm concentration (sperm / mL) in G-III, there was an increase (787.69 ± 480.72)compared to groups. It was concluded that the dose of 125mg / rbST increases the sperm concentration of ovineSanta Ines.(AU)

Efeito da melatonina sobre a integridade das membranas plasmática e mitocondrial de espermatozoides criopreservados de ovino / Effect of Melatonin on the plasmatic and mitochondrial integrity membrane of sperm cryopreserved ram

This study evaluate the effect of melatonin supplementation to TRIS-GEMA diluter in plasmatic andmitochondria membrane integrity of semen ram cryopreserved. Was used seven ejaculates of six ram,whichwere pooled in pool, according to the experimental groups: Control; T1 - 1x10-7 mol / L and T2 - 1x10-9 mol / Lmelatonin. Posteriorly the diluting the samples were packaged in straws of 0.25 ml and cryopreserved. Afterthawing at 37°C for 30 seconds, the samples were analyzed for Test thermoresistance slow (TTR) andacrosomal membrane integrity. For statistical analysis we used the Duncan test (α = 0.05) for meancomparison. Treatments 1 and 2 differed statistically control for plasma membrane integrity at times 60, 120and 180min. It was concluded that supplementation 1x10-7mol / L and 1x10-9mol / L of melatonin increased theintegrity of the plasma membrane to 60,120 and 180 minutes.(AU)

Caracterização física e estrutural de espermatozoides no ejaculado de Caprinos Azul, Canindé, Moxotó e Repartido / Physical and structural characterization of sperm in the ejaculate from Azul, Caninde, Moxotó and Repartido goat races

This study aimed to evaluate physically and structurally ejaculates from locally adapted goats in therainy season. Semen pool formed by the four goats of each breed was evaluated for physical, diluted in Tris-eggyolk and frozen. After thawing, it was diluted in Tris and incubated with carboxyfluorescein diacetate andpropidium iodide, to evaluate the integrity of the plasma membrane and JC1 to assess mitochondrial activity.The percentage of cells with damaged membrane and mitochondrial activity in spermatozoa between breed werecompared, every hour for three hours. There was no significant difference (P > 0,05) in relation to thepercentage of cells with damaged plasma membrane, as well as between the incubation times. The Azul breedhad lower mitochondrial activity (P < 0,05). The evaluated semen of all breeds showed no significant damage tothe plasma membrane in the rainy season, but the Azul breed had lower mitochondrial activity in sperm cells.(AU)

O inibidor de serino protease Antipaina preserva a integridade do acrossoma de espermatozoides bovino Curraleiro Pé-Duro durante a criopreservação / The inhibitor of serine protease Antipain preserve the integrity in the acrosome of sperm bovine Curraleiro Pé- Duro for cryopreservation

This study was conducted to evaluate the effect of different concentrations of inhibitor antipain serineprotease (10 g, 50 ug and 100 ug) in supplementing the dilutive of bovine semen freezing. Thirty-six ejaculatedfour Curraleiro Pé-Duro cattle were used for cryopreservation. The epifluorescence microscopy was used todetermine the acrosomal integrity and acrosome reaction rate was calculated after sperm were incubated for sixhours heparin. Spermatozoa cryopreserved in the presence of antipain significantly increased acrosomalintegrity, as they were able to complete the acrosome reaction in vitro. In conclusion, the inhibitor is antipainserine protease is able to preserve the integrity of cryopreserved sperm acrosomal cattle.(AU)

Efeito do Inibidor do ativador do plasminogênio 1 (PAI-1) nos parâmetros cinéticos de espermatozoides criopreservados de touros Curraleiro Pé Duro / Inhibitor effect of plasminogen activator 1 (PAI-1) in the kinetic parameters of sperm cryopreserved of Curraleiro Pé-Duro bulls

This study was conducted to evaluate the effect of different plasminogen activator inhibitor 1concentrations (70 ƞg, 140 and 210 ƞg ƞg) in the kinetic parameters of sperm cryopreserved of Curraleiro FootHard bulls. Sperm kinetics were analyzed by means of a computer aided system (CASA). The variables evaluatedwere: (MT-%) (MOP-um / s), (VCL - um / s), (VSL - um / s), (VAP-um / s), (LIN -%) ( Str-%) (ALH - uM)Wobble (WOB -%) and (BCF-Hz). The cryopreserved sperm in the presence of the inhibitor of plasminogenactivator 1 in a concentration of 210 ƞg decreased velocity parameters in a straight line (VSL - um / s) andlinearity (LIN -%). In conclusion, supplementation of the Inhibitor of plasminogen activator 1 (PAI-1),cryopreservation of bovine semen does not improve the kinetic parameters as compared to the control.(AU)

Captopril na produção in vitro de embriões bovinos? / Captopril in production in vitro of embryos bovine?

Objective to evaluate the effect of captopril in vitro production of bovine embryos. 472 COCs were usedfrom abattoir ovaries, which were screened, sorted and selected in grades I and II, and submitted to the PIVprocess. The IVM treatments were added in the following concentrations: T1 - control; T2 - 5 mM of captopril;T3 - 10 uM of captopril; and T4 - 15 captopril uM, the COCs were subjected to IVM, IVF CIV. The results showthat there was no statistically significant difference (P > 0.05) between treatments during maturation. Thevariables studied were cleavage and blastocyst rates. The captopril supplementation did not improve (P > 0.05)G1 cleavage rate - 61.84%; G2 - 71.00%; G3 - 68.87%; and G4 - 56.90%. Supplementation of 20μM ofcaptopril influenced the amount of viable embryos.(AU)

Efeito do captopril na maturação in vitro de oócitos bovinos / Effect of captopril in maturation in vitro of oocyte bovine

This study aimed to evaluate the effect of captopril on the in vitro maturation of oocytes. 1627 COCscattle slaughter houses were used, were subsequently screened, classified into grades I and II, according to themorphological quality, and submitted to the IVM process, distributed in four treatments: T1 - control; T2 - 5μMof captopril; T3 - 10μM of captopril; and T4 - 15μM of captopril, incubated in an incubator at 38.5 ° C with anatmosphere of 5% CO2. After 22 hours, the COCs were stripped to evaluate the rate of maturation, which wereas follows: T1 - 56.60% (120/212); T2 - 60.60% (240/396); T3 - 52.88% (174/329) and T4 - 58.30% (207/355).The results show that there was no statistically significant difference ( P> 0.05) between treatments duringmaturation, concluding that captopril supplementation did not affect the maturation of COCs.(AU)

Avaliação da integridade da membrana plasmática e do potencial de atividade mitocondrial de sêmen criopreservado de touros frente a ação do (Limoneno (R)-(+) / Evaluation of plasma membrane integrity and mitochondrial activity potential of cryopreserved semen of bulls on action of the Limonene (R) - (+)

This study was conducted to evaluate the effect of different concentrations of limonene (R)-(+) (50 µM,100 µM and 150 µM) in supplementing the diluter of bulls freezing semen. Thirty-six ejaculated from fourCurraleiro-Pé-Duro bulls were used for cryopreservation. The epifluorescence microscopy was used todetermine the plasmatic integrity and mitochondrial activity potential. No was observed effect on the integrity ofplasma membrane nor mitochondrial activity potential when cryopreserved by adding limonene R - (+). Theresults obtained in this study allow to conclude that supplementation limonene (R) - (+) on diluter of bullsfreezing semen does not interfere with the integrity of the plasma membrane or the potential of spermmitochondrial activity.(AU)

Ação do Limoneno (S)-(-) sobre a cinética espermática de sêmen criopreservado de touros / Action of the Limonene (S) - (-) on sperm kinetics of cryopreserved semen of bulls

This study was conducted to evaluate the effect of different concentrations of Limoneno (S)-(-) (50 µM,100 µM and 150 µM) in supplementing the diluter of bulls freezing semen. Thirty-six ejaculated from fourCurraleiro-Pé-Duro bulls were used for cryopreservation. The cryopreserved spermatozoa were submitted to postthawmotility sperm of computer assisted analysis (CASA) to evaluate the characteristics of spermatic kinetics.Observed that The different concentrations of limonene (S) - (-) did not affect the parameters of kinetics spermaticexcept for linearity (LIN). The addition of 150 μM limonene (S) - (-) significantly increased (P <0.05) the LINcompared to the control. The results obtained in the present study allow us to conclude that the supplementationof limonene (S) - (-) in cryopreservation bovine semen diluent did not interfere in kinetics sperm.

Avaliação da integridade da membrana plasmática e do potencial de atividade mitocondrial de sêmen criopreservado de touros frente a ação do (Limoneno (R)-(+) / Evaluation of plasma membrane integrity and mitochondrial activity potential of cryopreserved semen of bulls on action of the Limonene (R) - (+)

This study was conducted to evaluate the effect of different concentrations of limonene (R)-(+) (50 µM,100 µM and 150 µM) in supplementing the diluter of bulls freezing semen. Thirty-six ejaculated from fourCurraleiro-Pé-Duro bulls were used for cryopreservation. The epifluorescence microscopy was used todetermine the plasmatic integrity and mitochondrial activity potential. No was observed effect on the integrity ofplasma membrane nor mitochondrial activity potential when cryopreserved by adding limonene R - (+). Theresults obtained in this study allow to conclude that supplementation limonene (R) - (+) on diluter of bullsfreezing semen does not interfere with the integrity of the plasma membrane or the potential of spermmitochondrial activity.

Captopril na produção in vitro de embriões bovinos? / Captopril in production in vitro of embryos bovine?

Objective to evaluate the effect of captopril in vitro production of bovine embryos. 472 COCs were usedfrom abattoir ovaries, which were screened, sorted and selected in grades I and II, and submitted to the PIVprocess. The IVM treatments were added in the following concentrations: T1 - control; T2 - 5 mM of captopril;T3 - 10 uM of captopril; and T4 - 15 captopril uM, the COCs were subjected to IVM, IVF CIV. The results showthat there was no statistically significant difference (P > 0.05) between treatments during maturation. Thevariables studied were cleavage and blastocyst rates. The captopril supplementation did not improve (P > 0.05)G1 cleavage rate - 61.84%; G2 - 71.00%; G3 - 68.87%; and G4 - 56.90%. Supplementation of 20μM ofcaptopril influenced the amount of viable embryos.

Efeito do captopril na maturação in vitro de oócitos bovinos / Effect of captopril in maturation in vitro of oocyte bovine

This study aimed to evaluate the effect of captopril on the in vitro maturation of oocytes. 1627 COCscattle slaughter houses were used, were subsequently screened, classified into grades I and II, according to themorphological quality, and submitted to the IVM process, distributed in four treatments: T1 - control; T2 - 5μMof captopril; T3 - 10μM of captopril; and T4 - 15μM of captopril, incubated in an incubator at 38.5 ° C with anatmosphere of 5% CO2. After 22 hours, the COCs were stripped to evaluate the rate of maturation, which wereas follows: T1 - 56.60% (120/212); T2 - 60.60% (240/396); T3 - 52.88% (174/329) and T4 - 58.30% (207/355).The results show that there was no statistically significant difference ( P> 0.05) between treatments duringmaturation, concluding that captopril supplementation did not affect the maturation of COCs.

Efeito do Inibidor do ativador do plasminogênio 1 (PAI-1) nos parâmetros cinéticos de espermatozoides criopreservados de touros Curraleiro Pé Duro / Inhibitor effect of plasminogen activator 1 (PAI-1) in the kinetic parameters of sperm cryopreserved of Curraleiro Pé-Duro bulls

This study was conducted to evaluate the effect of different plasminogen activator inhibitor 1concentrations (70 ƞg, 140 and 210 ƞg ƞg) in the kinetic parameters of sperm cryopreserved of Curraleiro FootHard bulls. Sperm kinetics were analyzed by means of a computer aided system (CASA). The variables evaluatedwere: (MT-%) (MOP-um / s), (VCL - um / s), (VSL - um / s), (VAP-um / s), (LIN -%) ( Str-%) (ALH - uM)Wobble (WOB -%) and (BCF-Hz). The cryopreserved sperm in the presence of the inhibitor of plasminogenactivator 1 in a concentration of 210 ƞg decreased velocity parameters in a straight line (VSL - um / s) andlinearity (LIN -%). In conclusion, supplementation of the Inhibitor of plasminogen activator 1 (PAI-1),cryopreservation of bovine semen does not improve the kinetic parameters as compared to the control.

O inibidor de serino protease Antipaina preserva a integridade do acrossoma de espermatozoides bovino Curraleiro Pé-Duro durante a criopreservação / The inhibitor of serine protease Antipain preserve the integrity in the acrosome of sperm bovine Curraleiro Pé- Duro for cryopreservation

This study was conducted to evaluate the effect of different concentrations of inhibitor antipain serineprotease (10 g, 50 ug and 100 ug) in supplementing the dilutive of bovine semen freezing. Thirty-six ejaculatedfour Curraleiro Pé-Duro cattle were used for cryopreservation. The epifluorescence microscopy was used todetermine the acrosomal integrity and acrosome reaction rate was calculated after sperm were incubated for sixhours heparin. Spermatozoa cryopreserved in the presence of antipain significantly increased acrosomalintegrity, as they were able to complete the acrosome reaction in vitro. In conclusion, the inhibitor is antipainserine protease is able to preserve the integrity of cryopreserved sperm acrosomal cattle.