RESUMO

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.

Assuntos

RESUMO

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)

Assuntos

RESUMO

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)

Assuntos

RESUMO

O processo de fertilização é resultante de diversos mecanismos funcionais e estruturais do espermatozoide. Na andrologia veterinária, no que se refere à avaliação do potencial coeundi e generandi, estão disponíveis diversos testes laboratoriais e de imagem que servem como ferramenta de seleção de touros para a reprodução durante a estação de monta. Estas metodologias auxiliam nos protocolos de criopreservação espermática, na investigação de melhores crioprotetores e diluidores para uso in vivo ou in vitro. No entanto, são poucos utilizadas na rotina diária a campo e restritos aos centros de pesquisa. A presente revisão de literatura objetiva descrever as metodologias que estimam o potencial fecundante do sêmen criopreservado do touro.(AU)

The fertilization process is the result of several functional and structural mechanisms of the sperm. In veterinary andrology, with regard to the evaluation of the coeundi and generandi potential, several laboratory and image tests are available that serve as a tool for selecting bulls for breeding during the breeding season. These methodologies assist in sperm cryopreservation protocols, in the investigation of better cryoprotectants and extenders for use in vivo or in vitro. However, they are few used in the daily routine in the field and restricted to research centers. The present literature review aims to describe the methodologies that estimate the fertile potential of the bulls cryopreserved semen.(AU)

Assuntos

RESUMO

O processo de fertilização é resultante de diversos mecanismos funcionais e estruturais do espermatozoide. Na andrologia veterinária, no que se refere à avaliação do potencial coeundi e generandi, estão disponíveis diversos testes laboratoriais e de imagem que servem como ferramenta de seleção de touros para a reprodução durante a estação de monta. Estas metodologias auxiliam nos protocolos de criopreservação espermática, na investigação de melhores crioprotetores e diluidores para uso in vivo ou in vitro. No entanto, são poucos utilizadas na rotina diária a campo e restritos aos centros de pesquisa. A presente revisão de literatura objetiva descrever as metodologias que estimam o potencial fecundante do sêmen criopreservado do touro.

The fertilization process is the result of several functional and structural mechanisms of the sperm. In veterinary andrology, with regard to the evaluation of the coeundi and generandi potential, several laboratory and image tests are available that serve as a tool for selecting bulls for breeding during the breeding season. These methodologies assist in sperm cryopreservation protocols, in the investigation of better cryoprotectants and extenders for use in vivo or in vitro. However, they are few used in the daily routine in the field and restricted to research centers. The present literature review aims to describe the methodologies that estimate the fertile potential of the bulls cryopreserved semen.

Assuntos

RESUMO

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.

Assuntos

RESUMO

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.

Assuntos

RESUMO

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.

Assuntos

RESUMO

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.

Assuntos

RESUMO

A Leishmaniose Visceral (LV) é uma doença parasitária crônica, grave e endêmica em algumas regiões do Brasil, tendo os humanos e os cães como principais hospedeiros. Objetivou-se com este trabalho avaliar o perfil clínico, laboratorial e seminal de cães machos Sem Raça Definida (SRD) diagnosticados com LV, no município de Teresina, Piauí. Os animais foram provenientes da Gerência de Controle de Zoonoses do município e levados para o canil do Centro de Ciências Agrárias da Universidade Federal do Piauí, onde permaneceram de fevereiro a abril de 2014 sob avaliação e realização de exames. Foram coletadas amostras de sangue e sêmen de 12 cães, divididos em dois grupos, sendo 06 positivos (GI) e 06 negativos (GII) para a doença. Os resultados apontaram linfadenomegalia, emagrecimento e onicogrifose como principais sinais clínicos nos cães com LV e as alterações hematológicas foram, anemia normocítica normocrômica e trombocitopenia. A análise de bioquímica sérica revelou aumento nos índices de ureia nos animais parasitados no final do experimento, com diferença estatística entre os grupos, porém os níveis de creatinina mantiveram-se sem alteração. Quanto às análises seminais, foram encontradas diferenças significativas entre os grupos nos parâmetros de motilidade, vigor e concentração espermática, além da presença de patologias espermáticas que indicaram aumento de espermatozoides com alterações de cabeça e cauda. Conclui-se que cães com LV apresentam alterações clínicas, laboratoriais e seminais que podem prejudicar a condição geral e reprodutiva dessa espécie animal.

The Visceral Leishmaniasis (VL) is a chronic and severe parasitic disease, endemic in some regions of Brazil, and having dogs and humans as the main hosts. The objective of this work was to evaluate the clinical, laboratory and seminal profile of Mixed breed male dogs diagnosed with VL, in the city of Teresina, Piauí. The animals were obta ined from the Zoonoses Control Management of city and taken to the kennel of the Agrarian Sciences Center of the Federal University of Piauí, where they remained from February to April 2014 under evaluation and exams. Blood and semen samples were collected from 12 dogs, divided into two groups, being 06 positive (GI) and 06 negative (GII) for the disease. The results indicated lymphadenomegaly, weight loss and onychogryphosis as the main clinical signs in dogs with VL and the hematological changes were normochromic normocytic anemia and thrombocytopenia. The analysis of serum biochemistry revealed an increase in urea indexes in the parasitized animals at the end of the experiment, with statistical difference between the groups, however the levels of creatinine remained unchanged. Regarding the seminal analyzes, significant differences were found between the groups in the parameters of motility, vigor and sperm concentration, in addition to the presence of sperm pathologies that indicated an increase in sperm with changes of the head and tail. It is concluded that dogs with VL present clinical, laboratory and seminal alterations that can impair the general and reproductive condition of this animal species.

Assuntos