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.

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)

Raiva em equino na região Sul do estado do Piauí / Equine rabies in the Southern region of Piauí state

Background: Rabies is an infectious disease that is important in the “One Health” worldwide with high lethality rate. The etiological agent is a neurotropic virus, genus Lyssavirus, transmitted mainly through the saliva of infected animals. For equines, the bite of hematophagous bats is the main source of infection. Piauí is an important state for equestrian sports and the increase in the number of horses with neurological clinical signs without diagnosis has increased in recent years. In this context, the aim of this study is to report to the scientific community a confirmed case of equine rabies in the Santa Luz county, Southernmost state of Piauí, Brazil. Case: A 3-year-old female non-defined breed horse, was admitted to the Hospital Veterinário da Universidade Federal do Piauí (UFPI/CPCE). The equine had difficulty walking 2 days ago, in the panoramic inspection was restless and disoriented in the paddock. Rectal temperature of 38.2°C, heart rate of 60 bpm, respiratory rate of 40 mpm, congested mucosa and dyspnea were verified. With the progression of the neurological signals, it positioned itself in a lateral decubitus with pedaling movements, hyperesthesia, dysphagia and paralysis of the hindlimbs. The clinical suspicion was rabies and the Agência de Defesa Agropecuária do Piauí (ADAPI) was communicated to euthanize the animal and collect samples for diagnosis in accordance with official standards of the Ministério da Agricultura, Pecuária e Abastecimento (MAPA). At necropsy, there was slight brain hyperemia, with no other significant organ changes. Fragments of the cerebellum, cortex, hippocampus and spinal cord were collected and sent at a temperature of 4°C to perform the Direct Immunofluorescence (DIF) assay. Samples for histopathology were not collected because they do not include assay for confirmatory diagnosis of rabies. The DIF technique with...

Raiva em equino na região Sul do estado do Piauí / Equine rabies in the Southern region of Piauí state

Background: Rabies is an infectious disease that is important in the “One Health” worldwide with high lethality rate. The etiological agent is a neurotropic virus, genus Lyssavirus, transmitted mainly through the saliva of infected animals. For equines, the bite of hematophagous bats is the main source of infection. Piauí is an important state for equestrian sports and the increase in the number of horses with neurological clinical signs without diagnosis has increased in recent years. In this context, the aim of this study is to report to the scientific community a confirmed case of equine rabies in the Santa Luz county, Southernmost state of Piauí, Brazil. Case: A 3-year-old female non-defined breed horse, was admitted to the Hospital Veterinário da Universidade Federal do Piauí (UFPI/CPCE). The equine had difficulty walking 2 days ago, in the panoramic inspection was restless and disoriented in the paddock. Rectal temperature of 38.2°C, heart rate of 60 bpm, respiratory rate of 40 mpm, congested mucosa and dyspnea were verified. With the progression of the neurological signals, it positioned itself in a lateral decubitus with pedaling movements, hyperesthesia, dysphagia and paralysis of the hindlimbs. The clinical suspicion was rabies and the Agência de Defesa Agropecuária do Piauí (ADAPI) was communicated to euthanize the animal and collect samples for diagnosis in accordance with official standards of the Ministério da Agricultura, Pecuária e Abastecimento (MAPA). At necropsy, there was slight brain hyperemia, with no other significant organ changes. Fragments of the cerebellum, cortex, hippocampus and spinal cord were collected and sent at a temperature of 4°C to perform the Direct Immunofluorescence (DIF) assay. Samples for histopathology were not collected because they do not include assay for confirmatory diagnosis of rabies. The DIF technique with...(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)

Maturação de oócitos bovinos em meio suplementado com captopril / Maturation of Bovine oocytes in medium supplemented with captopril

The aim of this research was to evaluate the effects of supplementation of maturation medium withcaptopril in oocyte in vitro maturation (IVM). 470 CCOs were recovered and classified into grades I and II,being divided into four groups: G1 (n = 56), that was control group; G2 (n = 152) 20 mM of captopril; G3 (n =126) 40 uM of captopril; and G4 (n = 136) 80 uM of captopril, then these were subsequently submitted to IVMprocess for 24 hours at 38.5°C in 5% CO2. After 24 hours of maturation, oocytes were denuded and evaluatedfor the first polar body extrusion and were, therefore, considered matured. The addition of the Captopril inmedium of oocytes maturation has not improved the IVM rate.(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)

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)

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 do efeito da melatonina sobre o grau de qualidade de embriões bovinos produzidos in vitro / Evaluation of the effect of melatonin on the quality grade of bovine embryos produced in vitro

The present study evaluated the effect of melatonin on the degree of embryonic quality during theprocess of in vitro production of embryos. 512 cumulus-oocyte complexes of type I and II were submitted to invitro production of embryos with 0 and 100 µM of melatonin in the maturation medium. There was no differencerelative to the rate of cleavage 36.44% vs 32.93% and total viable embryos 16.44% vs 12.16% (P <0.05).Likewise, there was no difference in the degree of embryonic quality (P <0.05). Therefore, the mediumsupplemented with melatonin was not able to improve the cleavage rate, total viable embryos and quality ofembryos fertilized in vitro.(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.(AU)

Produção in vitro de embriões bovinos em meio suplementado com captopril / In vitro production of embryos Cattle in medium supplemented with captopril

The aim of this study was to evaluate the effects of supplementation of maturation medium withcaptopril, and its consequences in the bovine maturation, fertilization and embryo development in vitro. 326bovine ovaries from a local slaughterhouse were used. 1101 CCOs were recovered and distributed in fourgroups: G1 (n = 112), that was control group; G2 (n = 322) 20μM of captopril; G3 (n = 367) 40μM ofcaptopril; and G4 (n = 300) captopril 80μM and later submitted to IVM. The matured CCOs were fertilized, andco-incubated. After fertilization, 676 presumptive zygotes were cultured and maintained in the greenhouse for 7days. The total number of viable embryos was 12; 39; 32 and 31, respectively in the experimental groups.Considering the experimental conditions adopted, it was concluded that the addition of the Captopril in mediumof oocytes IVM positively doesnt influence embryonic development, as evidenced by the similar percentages ofembryo production.(AU)

Histopatologia e Imunohistoquímica de testículos de cães naturalmente infectados por Leishmania sp / Histopathology and Immunohistochemistry testicles of dogs naturally infected by Leishmania sp

The objective of this study was to evaluate testicular changes and the presence of Leishmania sp. intesticles of dogs with Visceral Leishmaniasis (LV). The animals were obtained from Teresina Zoonosis ControlAdministration, and taken to the kennels of Agricultural Sciences Center of the Federal University of Piauí(UFPI), where they stayed for two months and were subsequently euthanized for removal of the testicles. 12 dogswere used, 06 positive (GI) and 06 negative (GII) for LV and conducted examinations of histopathology andimmunohistochemistry (IMH) of the testicles. All were negative by IMH technique and by histopathology. In thisstudy, we observed testicular lesions in all animals, with no direct relationship between the presence ofLeishmania sp. and intensity of the damage found in the reproductive system, reducing the possibility of venerealtransmission between oligositomatics dogs.(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.

Avaliação do efeito da melatonina sobre o grau de qualidade de embriões bovinos produzidos in vitro / Evaluation of the effect of melatonin on the quality grade of bovine embryos produced in vitro

The present study evaluated the effect of melatonin on the degree of embryonic quality during theprocess of in vitro production of embryos. 512 cumulus-oocyte complexes of type I and II were submitted to invitro production of embryos with 0 and 100 µM of melatonin in the maturation medium. There was no differencerelative to the rate of cleavage 36.44% vs 32.93% and total viable embryos 16.44% vs 12.16% (P <0.05).Likewise, there was no difference in the degree of embryonic quality (P <0.05). Therefore, the mediumsupplemented with melatonin was not able to improve the cleavage rate, total viable embryos and quality ofembryos fertilized in vitro.

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.