Efeito da adição do ácido palmítico e da vitamina E ao diluidor Tris-gema na criopreservação de sêmen canino / Effect of addition of palmitic acid and Vitamin E to the Tris-egg yolk diluter in canine semen cryopreservation

Canine sperm is a very delicate cell that is quite susceptible to oxidative stress since the cytoplasm is restricted and features little antioxidant reserves. Furthermore, the sperm membrane has some polyunsaturated fatty acids sensitive to lipid peroxidation, which makes it important to addition antioxidant substances to the diluter aiming at decreasing such stress to the sperm cell, particularly during seminal cryopreservation. Several antioxidants have been used in this process in some domestic animal's species, however, the use of palmitic acid has been little reported in works on cryopreservation of semen of the canine species. Hence, this study aimed to assess the effect of addition antioxidants palmitic acid and vitamin E to the Tris-egg yolk diluter on the semen quality of dogs after thawing. Samples were collected from the ejaculates of 4 adult dogs, apparently healthy, of the American Pit Bull Terrier breed of kennels in the city of Teresina, PI, places where the pre-freezing procedures of the dog's semen were performed. The samples were diluted in Tris citric acid fructose (3.28 g Tris-hydroxymethyl-aminomethane, 1.78 g citric acid monohydrate and 1.25 g D-fructose), dissolved in 100 mL distilled water, and added 20% egg yolk and 6% glycerol, at the concentration of 100x106 sptz/mL. The semen samples were divided into 3 mL aliquots to form 3 experimental groups: G1 - Only Tris-egg yolk (Control group); G2 - Tris-egg yolk + 100 µM palmitic acid; and G3 - Tris-egg yolk + 116 µM vitamin E. Semen was collected weekly over a period of little over 2 months. After thawing, thermorresistance test (TTR) was carried out at 0, 30, 60, and 90 min to assess spermatics motility and vigor, in addition to analysis of integrity of plasma membrane, acrosomal membrane and mitochondrial activity of the sperm, using fluorescent probes. These assessments were performed out at the Animal Reproduction Biotechnology Laboratory (LBRA/UFPI). In the TTR, G2 and G3 didn't exhibit significant results for spermatics motility or vigor when compared with the control group. The palmitic acid and vitamin E also had no significant effects on the parameters of acrosomal membrane integrity or mitochondrial activity. However, sperm cryopreserved with the addition of palmitic acid exhibited significant differences for plasma membrane integrity, providing greater protection to the sperm cells in G2. The palmitic acid is one of the most saturated fatty acids in human semen, with reports of great proportions also in the seminal plasma of dogs. Its main role is to protect the plasma membrane from external damage, improving viability and fertility of the sperm after cryopreservation. Data is scarce in the literature on the composition of fatty acids in canine semen and regarding the use of palmitic acid as a seminal antioxidant in that species, which grants further studies aiming to investigate such valuable information for canine reproduction. It is concluded that addition palmitic acid at 100µM concentration to the Tris-egg yolk diluter was able to preserve the integrity of the plasma membrane during the process of cryopreservation of canine semen.(AU)

Análises da membrana plasmática, acrossoma e mitocôndrica de espermatozoide ovino após criopreservação com adição de Ácido Fólico no meio tris-gema / Plasma membrane, acrosome and mitochondria analysis of sheep sperm after cryopreservation with addition of folic acid in the tris-yolk medium

Folic acid is closely linked to cobalamin, which is a carrier of hydroxymethyl and ant groups. The objective of this work was to evaluate the effect of adding folic acid to the TRIS-yolk diluter and possible effect on the membrane, mitochondria and acrosome of sheep sperm after thawing the semen. There were six sheep and seven collections of each animal in sessions between 48 and 72 hours. After collection and analysis, the semen samples were mixed and submitted to the formation of a pool. Diluted in TRIS-yolk medium, and divided into 3 groups; Group control; Group 2: 10,000 µM of folic acid and in Group 3: 5000 µM of folic acid. Subsequently, the semen samples were packaged in 0.25 mL straws and processed in a semen freezing machine. After a few days, the semen was thawed and evaluated: plasma membrane integrity, acrosome function and integrity. They were analyzed using an analysis of variance (ANOVA) followed by the Newman-Keuls test, using SAS. The additionof a micronutrient with potential antioxidant action in the semen indicates that it can reduce the action of free radicals that alter the plasma membrane and sperm DNA. The evaluation of the plasma membrane integrity, mitochondrial activity and the acrosome integrity of post-thawed sperm were not significantly different between the groups evaluated. Thus, it is concluded that the addition of folic acid in concentrations of 5000 µM and 10000 µM to the TRIS-yolk seminal diluter does not significantly influence the variables evaluated in this experiment.

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.

Análises de membrana plasmática, acrossoma e mitocôndria de espermatozoides ovino após criopreservação com adição do ácido fólico no meio Tris-gema / Plasma membrane, acrosome and mitochondria analysis of sheep sperm after cryopreservation with addition of folic acid in the tris-yolk medium

Folic acid is closely linked to cobalamin, which is a carrier of hydroxymethyl and ant groups. The objective of this work was to evaluate the effect of adding folic acid to the TRIS-yolk diluter and possible effect on the membrane, mitochondria and acrosome of sheep sperm after thawing the semen. There were six sheep and seven collections of each animal in sessions between 48 and 72 hours. After collection and analysis, the semen samples were mixed and submitted to the formation of a pool. Diluted in TRIS-yolk medium and divided into 3 groups; Group control; Group 2: 10,000µM of folic acid and in Group 3: 5000µM of folic acid. Subsequently, the semen samples were packaged in 0.25mL straws and processed in a semen freezing machine. After a few days, the semen was thawed and evaluated: plasma membrane integrity, acrosome function and integrity. They were analyzed using an analysis of variance (ANOVA) followed by the Newman-Keuls test, using SAS. The additionof a micronutrient with potential antioxidant action in the semen indicates that it can reduce the action of free radicals that alter the plasma membrane and sperm DNA. The evaluation of the plasma membrane integrity, mitochondrial activity and the acrosome integrity of post-thawed sperm were not significantly different between the groups evaluated. Thus, it is concluded that the addition of folic acid in concentrations of 5000µM and 10000µM to the TRIS-yolk seminal diluter does not significantly influence the variables evaluated in this experiment.

Análises da membrana plasmática, acrossoma e mitocôndrica de espermatozoide ovino após criopreservação com adição de Ácido Fólico no meio tris-gema / Plasma membrane, acrosome and mitochondria analysis of sheep sperm after cryopreservation with addition of folic acid in the tris-yolk medium

Folic acid is closely linked to cobalamin, which is a carrier of hydroxymethyl and ant groups. The objective of this work was to evaluate the effect of adding folic acid to the TRIS-yolk diluter and possible effect on the membrane, mitochondria and acrosome of sheep sperm after thawing the semen. There were six sheep and seven collections of each animal in sessions between 48 and 72 hours. After collection and analysis, the semen samples were mixed and submitted to the formation of a pool. Diluted in TRIS-yolk medium, and divided into 3 groups; Group control; Group 2: 10,000 µM of folic acid and in Group 3: 5000 µM of folic acid. Subsequently, the semen samples were packaged in 0.25 mL straws and processed in a semen freezing machine. After a few days, the semen was thawed and evaluated: plasma membrane integrity, acrosome function and integrity. They were analyzed using an analysis of variance (ANOVA) followed by the Newman-Keuls test, using SAS. The additionof a micronutrient with potential antioxidant action in the semen indicates that it can reduce the action of free radicals that alter the plasma membrane and sperm DNA. The evaluation of the plasma membrane integrity, mitochondrial activity and the acrosome integrity of post-thawed sperm were not significantly different between the groups evaluated. Thus, it is concluded that the addition of folic acid in concentrations of 5000 µM and 10000 µM to the TRIS-yolk seminal diluter does not significantly influence the variables evaluated in this experiment.(AU)

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)

Análises de membrana plasmática, acrossoma e mitocôndria de espermatozoides ovino após criopreservação com adição do ácido fólico no meio Tris-gema / Plasma membrane, acrosome and mitochondria analysis of sheep sperm after cryopreservation with addition of folic acid in the tris-yolk medium

Folic acid is closely linked to cobalamin, which is a carrier of hydroxymethyl and ant groups. The objective of this work was to evaluate the effect of adding folic acid to the TRIS-yolk diluter and possible effect on the membrane, mitochondria and acrosome of sheep sperm after thawing the semen. There were six sheep and seven collections of each animal in sessions between 48 and 72 hours. After collection and analysis, the semen samples were mixed and submitted to the formation of a pool. Diluted in TRIS-yolk medium and divided into 3 groups; Group control; Group 2: 10,000µM of folic acid and in Group 3: 5000µM of folic acid. Subsequently, the semen samples were packaged in 0.25mL straws and processed in a semen freezing machine. After a few days, the semen was thawed and evaluated: plasma membrane integrity, acrosome function and integrity. They were analyzed using an analysis of variance (ANOVA) followed by the Newman-Keuls test, using SAS. The additionof a micronutrient with potential antioxidant action in the semen indicates that it can reduce the action of free radicals that alter the plasma membrane and sperm DNA. The evaluation of the plasma membrane integrity, mitochondrial activity and the acrosome integrity of post-thawed sperm were not significantly different between the groups evaluated. Thus, it is concluded that the addition of folic acid in concentrations of 5000µM and 10000µM to the TRIS-yolk seminal diluter does not significantly influence the variables evaluated in this experiment.(AU)

Ação do extrato etanólico de Mufumbo sobre a criopreservação do sêmen de garanhão / Action of the ethanol extract of Mufumbo on cryopreservation of stallion sêmen

Various natural antioxidants are being added to the freezing diluents in order to minimize deleteriouseffects to the membrane of spermatozoa during the cryopreservation process. Avaliou if stallions semencharacteristics cryopreserved amid thinner Botu-crio plus ethanol extract from the bark of mufumbo(Combretum leprosum) at different concentrations (30 mg, 60 mg and 120 mg). Samples of each treatment weresubmitted to hypoosmotic test membrane integrity tests and mitochondrial function (fluorescent tubes); Therewas no significant difference (P > 0.05) and the percentage of spermatozoa reactive to hiposmotic test betweenthe groups tested. The percentage of sperm with intact plasma membrane, as well as the mitochondrial potentialwere not statistically different (P > 0.05) between the control and treatment.(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)

Efeito da Somatrotopina Bovina Recombinante (rbST) associado a um protocolo de Inseminação Artificial em Tempo Fixo em cabras SPRD / Effect of Recombinant Bovine Somatotropin (rbST) associated with a protocol Artificial Insemination in Fixed Time in SPRD goats

We evaluated the effect of Recombinant Bovine Somatotropin (rbST) seven days before the start of thesynchronization protocol on pregnancy rate in goats. Were used 101 females without defined breed in semiintensivesystem, randomly divided into two groups. Seven days before the start of estrus synchronization, G1 (n= 49) received 125 mg of rbST, IM, and G2 (n = 52) physiological saline, subcutaneously. In D0, the groupsreceived 1 mL of BE, IM, and were synchronized with intravaginal sponges impregnated with 60 mg ofmedroxyprogesterone for 11 days. In D9, they received 300 IU of eCG and 75 µg of PGF2α. On D11, thesponges were removed, we performed IATF 36 and 48 hours later, transcervically. It was used pool of AngloNubians semen. Gestational diagnosis was performed 30 days later. A single dose administration of rbST, sevendays before estrus synchronization, did not increase the pregnancy rate.(AU)

Efeito da melatonina sobre a viabilidade de espermatozóides criopreservados de ovinos / Effect of melatonin on the feasibility of sperm cryopreserved of ram

This study to evaluate the effect of melatonin supplementation to TRIS-GEMA diluter in sperm viabilityof semen ram cryopreserved. Was used seven ejaculates of six ram,which were pooled in pool, according to theexperimental groups: T1 - control; T2 - 1x10-7 mol / L and T3 - 1x10-9 mol / L melatonin. Posteriorly thediluting the samples were packaged in straws of 0.25 ml and cryopreserved. After thawing at 37°C for 30seconds, the samples were analyzed for Test thermoresistance slow (TTR) and acrosomal membrane integrity.For statistical analysis we used the Duncan test (α = 0.05) for mean comparison. The results demonstrated thatto the TTR at time 0 min, T2 showed 54.28% motility, and T1obtained lower values in the times 0 and 60 minutesfor force. Supplementation of 1x10-9mol / L of melatonin improved total motility.(AU)

Avaliação do chá do extrato de copaíba (Copaifera luetzelburgii) pós-criopreservação de sêmen de caprinos pelo teste de termoresistência / Evaluation of the tea of copaíba extract (Copaifera luetzelburgii) in goat semen post-thawing by term-resistance test

The semen cryopreservation process causes damages in spermatozoa characteristics. This study aimedto evaluate the effects of tea of Copaíba extract (Copaifera luetzelburgii), in different concentrations, added tothe Tris-egg yolk extender, in the quality of post-cryopreserved spermatozoa from locally adapted breeds ofgoats. Were used four (4) goats Breeds (Azul, Canindé, Moxotó e Repartido) adults, male, six of each breed,aged between 1 and 6 years, a pool of ejaculates were formed to analyze. The animals were randomly dividedinto four experimental treatments: TI (Control), TII (0,1mg), TIII (0,2mg) and TIV (0,5mg). The samples werefrozen and stored, after 30 days they were thawed for analysis by term-resistance test (TTR). The addition of teaof copaíba extract showed no significant difference between treatments by TTR, therefore the use of tea copaíbaextract in seminal diluting not increment the sperm longevity post-thawing.(AU)

Adição do Ácido Fólico na criopreservação de sêmen ovino da raça Santa Inês / Addition of Folic Acid in the cryopreservation of semen from Santa Ines sheep

We evaluated the addition of folic acid to the extender TRIS-egg yolk used in cryopreservation, seekingimprovements for the sperm kinetics and mitochondrial activity post-thawing. Were evaluated sevenejaculates/animal from six Santa Inês by Artificial Vagina, with minimum values of motility by 70.0% and vigor3. We evaluated the seminal pool for concentration and morphology. We diluted in Tris-egg yolk extender forcryopreservation, dividing it in control, Group 2 we added 10.000 uM and Group 3 we added 5000 uM of folicacid. We froze in straws(0.25 mL). Was evaluated for motility and vigor, post-thawing by slow term-resistancetest, the times T0, T60, T120 and T180 minutes one week after. For the mitochondrial activity evaluation, wasused cationic lipophilic fluorochrome JC-1. The folic acid effects were analyzed using ANOVA followed byStudent Newman-Keuls test. The groups with folic acid showed significant higher motility. Mitochondrialactivity did not differ between groups.(AU)

Avaliação morfológica, funcional e tempo de termorresistência de espermatozoides caprinos localmente adaptados / Morphological, functional and thermoresistance time evaluation of locally adapted goats sperm

This study evaluated the quality of ejaculated from adapted breeds of goats as the morphologic aspects,functional and thermoresistance. An aliquot of the pool of each breed was used for morphological evaluation bywet preparation, quantifying sperm defects and dividing them into classes. The samples were cryopreservedusing TK3000 machine and evaluated by thermoresistance test by checking motility and sperm vigor every hour.For plasma membrane functionality analysis, was used hiposmotic test. There was no difference (P > 0.05)between the breeds in all sperm defects before and after cryopreservation, as well as for the thermoresistancetimes for motility and sperm vigor (P > 0.05). There was a significant reduction in motility and sperm vigor 120minutes after thawing of the samples. There was no difference (P > 0.05) between the percentage of sperm withfunctional membrane between the evaluated breeds. Therefore, the evaluated breeds have morphology,functionality and similar sperm thermoresistance.(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)

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.

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.

Eficiência da inseminação artificial em tempo fixo (IATF) associada à detecção de estro por marcação da cauda e presença de muco vaginal no dia da inseminação em vacas Nelore / Efficiency of artificial inseminatio n in fixed time (FTAI) associated with estrus detection by marking the tail and the presence ofvaginal mucus on the day of insemination in Nelore cows

The objective of this study was to evaluate the efficiency of artificial insemination in fixed timeassociated with estrus detection in Nellore cows. They used 917 females, all submitted to estrus synchronizationprotocol and formed two groups: G1 (n = 489), marked on the back with fluorescent dye and then checked if thecroup was blurred or not, and had vaginal mucus or not and G2 (n = 428) were formed by females inseminatedonly synchronized. Pregnancy diagnosis was performed by ultrasound 30 days after insemination. In G1 and G2,pregnancy rates were, respectively, 46.01% and 41.12%. In G1, blurred females (52.19%), not blurred(21.78%), presence of mucus (62.57%) and no mucus (35.97%). Pregnancy rates did not differ between thegroups G1 and G2. In G1, blurred females or mucus had higher rates. It is concluded that the detection ofestrus, insemination favored the pregnancy rate.