The aim of this research was to evaluate the use of centrifuged egg yolk (CEY) in the freezing process of canine epididymal spermatozoa. Testicular-epididymis complexes (TECs) were obtained after the orchiectomy of 20 domestic dogs (Canis familiaris) of different breeds and with an age range between 1 and 6 years. The tail of the epididymis was dissected and spermatozoa were obtained by the retrograde flow technique. The spermatozoa were divided into two aliquots and extended in Triladyl® supplemented with 10% CEY (CEY10) or 20% CEY (CEY20). Semen freezing was performed using a standard protocol with liquid nitrogen vapors and semen was packed in straws of 0.5 ml. A computerized system was used to evaluate the motility, kinetics and concentration of spermatozoa. In addition, post-thawing sperm vitality (VE), abnormal morphology (MA) and plasma membrane integrity (HOS) were evaluated. St...

Isoespintanol from the leaves of Oxandra cf xylopioides and thymol present in the essential oils of thyme (Thymus vulgaris L) and oregano (Origanum vulgare L) is known for their activity as antioxidants in different media. In this study, the antioxidant activity of isoespintanol and thymol in cryopreserved stallion semen was evaluated. The semen of five Colombian Creole horses was collected and subjected to a freezing protocol, in an extender supplemented with isoespintanol or thymol. The production of reactive oxygen species and the lipid peroxidation of the thawed semen were evaluated. As a result, a post-thawing reduction of the production of reactive oxygen species of semen was observed due to the effect of both antioxidants, whereas no difference was found in the lipid peroxidation levels of post-thawed semen. Isoespintanol and thymol act as antioxidants in stallion semen subjected ...

The seminal plasma (SP) has enzymatic and non-enzymatic antioxidants with the function of protecting sperm from the harmful effects of oxidative stress. The aim of this study was to evaluate the antioxidant contribution of SP during the freezing of equine semen and its effect on post-thawing sperm quality. The semen of five horses was collected, the SP was separated and supplemented in proportions of 0% (Control), 10% (PS10) and 20% (PS20) to the sperm fraction in the freezing extender. The total antioxidant capacity (TAC) was evaluated by the oxygen radical trapping capacity (ORAC) and ferric reducing capacity (FRAP) methods, and the activity of SOD, GPx and catalase enzymes by spectrophotometry. Fifteen ejaculates were subjected to freezing and motility (MOT), vitality (EV), morphology (AM), functional membrane integrity (MI) and mitochondrial membrane potential (MMP) of the sperm were...

Oxidative stress has been associated with the development of many chronic and degenerative diseases in canines. The presence of antioxidants in food contributes to the reduction of the harmful effects of reactive oxygen species (ROS) and nitrogen (RNS). Polyphenols are a group of substances with high antioxidant activity and play an important role in the prevention of diseases. The aim of this study was to evaluate the relationship between the composition, the antioxidant capacity and the polyphenol content of dog feeds from different commercial segments. Nineteen dog foods from four commercial segments: popular, economic, premium and super premium were randomly selected. Through a bromatological analysis the composition of each feed was evaluated. Total antioxidant capacity (TAC) and total phenolic content (TFC) were measured through the ABTS and Folin-Ciocalteu methods, respectively. A...

El objetivo de esta investigación fue evaluar el uso de yema de huevo centrifugada (YHC) en la congelación de espermatozoides epididimales caninos. Se obtuvieron los complejos testículo-epidídimo (CTE) después de la orquiectomía de 20 perros domésticos (Canis familiaris) de varias razas y en un rango de edad entre 1 y 6 años. Se diseccionó la cola del epidídimo y se obtuvieron los espermatozoides mediante la técnica de flujo retrógrado. Los espermatozoides obtenidos se dividieron en dos alícuotas y se diluyeron en Triladyl® suplementado con YHC al 10% (YHC10) o YHC al 20% (YHC20). Se realizó la congelación del semen mediante un protocolo convencional con vapores de nitrógeno líquido en pajillas para 0.5 ml. Se utilizó un sistema computarizado para evaluar la movilidad, la cinética y la concentración de los espermatozoides. Adicionalmente, se evaluaron posdescongelaci...

El isoespintanol proveniente de las hojas de Oxandra cf xylopioides y el timol presente en los aceites esenciales del tomillo (Thymus vulgaris L) y el orégano (Origanum vulgare L) son conocidos por su actividad antioxidante en diferentes medios. En este estudio se evaluó la actividad antioxidante del isoespintanol y el timol en el semen equino criopreservado. Se colectó el semen de cinco caballos criollos colombianos y se sometió a un protocolo de congelación, en un diluyente suplementado con isoespintanol o timol. Se evaluó la producción de especies reactivas de oxígeno y la peroxidación lipídica del semen descongelado. Como resultado, se observó una reducción pos-descongelación de la producción de especies reactivas de oxígeno del semen por efecto de ambos antioxidantes, mientras no se encontró diferencia entre los niveles de peroxidación lipídica del semen pos-descon...

The seminal plasma (SP) has enzymatic and non-enzymatic antioxidants with the function of protecting sperm from the harmful effects of oxidative stress. The aim of this study was to evaluate the antioxidant contribution of SP during the freezing of equine semen and its effect on post-thawing sperm quality. The semen of five horses was collected, the SP was separated and supplemented in proportions of 0% (Control), 10% (PS10) and 20% (PS20) to the sperm fraction in the freezing extender. The total antioxidant capacity (TAC) was evaluated by the oxygen radical trapping capacity (ORAC) and ferric reducing capacity (FRAP) methods, and the activity of SOD, GPx and catalase enzymes by spectrophotometry. Fifteen ejaculates were subjected to freezing and motility (MOT), vitality (EV), morphology (AM), functional membrane integrity (MI) and mitochondrial membrane potential (MMP) of the sperm were...

Oxidative stress has been associated with the development of many chronic and degenerative diseases in canines. The presence of antioxidants in food contributes to the reduction of the harmful effects of reactive oxygen species (ROS) and nitrogen (RNS). Polyphenols are a group of substances with high antioxidant activity and play an important role in the prevention of diseases. The aim of this study was to evaluate the relationship between the composition, the antioxidant capacity and the polyphenol content of dog feeds from different commercial segments. Nineteen dog foods from four commercial segments: popular, economic, premium and super premium were randomly selected. Through a bromatological analysis the composition of each feed was evaluated. Total antioxidant capacity (TAC) and total phenolic content (TFC) were measured through the ABTS and Folin-Ciocalteu methods, respectively. A...

Cuniculus paca is a terrestrial rodent that inhabits the neotropical forests of Central and South America. The aim of this study was to evaluate the effect of sperm separation on the quality of cryopreserved semen of C. paca. Sixty straws of frozen semen were thawed. One aliquot was subjected to sperm separation by single-layer density gradient centrifugation (SLC) and the other aliquot remained as a control group with semen not separated. Sperm motility and kinetics were evaluated using a computerized-assisted sperm analysis (CASA) system, functional membrane integrity by hypoosmotic test (HOST), and sperm morphology by eosin-nigrosin staining. The adjustment of linear models and comparison of means by Tukey was performed. The SLC separation produced a higher proportion of motile and linear spermatozoa, while the control showed higher values for curvilinear velocity (p˂0.05). Likewise,...

The aim of this study was to evaluate sodium caseinate (CS) in the dilution and freezing of bovine semen. Fifteen ejaculates from five bulls were diluted and then individually frozen using a diluent composed of tris-glycerol-citric acid with 8% v/v of low-density lipoproteins (LDL) and supplemented with CS in four treatments: CS 3% w/v (CS3), CS 4% w/v (CS4), CS 5% w/v (CS5) and without CS (Control). The mobility and kinetics (SCA® System) and the functional integrity of the membrane (HOS test) of the fresh, extended and thawed semen were evaluated. In addition, the morphology and vitality (eosin-nigrosin) of the fresh and extended semen, and the structural integrity of the membrane (SYBR14/IP), the acrosome integrity (FITC-PNA) and the in vitro fertilizing capacity (IVF) of thawed semen were evaluated. The statistical analysis of the sperm quality parameters was performed using linear ...

Se desconoce en gran medida el aporte del plasma seminal a la criotolerancia del semen equino. Algunos componentes del plasma seminal son conocidos por su acción antioxidante y, en consecuencia, podrían influir en la estabilidad oxidativa de los lípidos y las proteínas del semen. Esta investigación tuvo como objetivo evaluar la relación entre la estabilidad oxidativa y algunos componentes del plasma seminal con la criotolerancia del semen equino. En el plasma seminal de 30 caballos se evaluó el contenido de vitamina C (VC) por HPLC, proteínas totales (PT) por Bradford, y la oxidación proteica (OP) y lipídica (PL) por medición de carbonilos y el ensayo TBARS, respectivamente. El semen se congeló con 10% de plasma seminal y se evaluó la calidad seminal posdescongelación. Se realizó el ajuste de modelos lineales generalizados (GLM), análisis de regresión y correlación, y l...

Extenders have been established as an alternative to compensate the antioxidant deficiencies of stallion semen during freezing. Several research works have aimed to evaluate the effect of various additives with antioxidant properties on the equine sperm cryotolerance; however, in limited occasions this has been linked with the extender total antioxidant capacity (TAC). This research aimed to evaluate the effect of two additives on freezing extender TAC and stallion semen cryotolerance. Semen of five Colombian Creole horses was frozen in an extender under three treatments: T0: control, T1: quercetin 100 μM, T2: phosphates and carbonates (Na2HPO4 0.32 mg/ml + KH2PO4 0.67 mg/ml + K2CO3 0.39 mg/ml). TAC of extender was evaluated by the ABTS test. Post-thaw motility and kinetics, abnormal morphology (AM), and structural (MSI) and functional (HOS) plasma membrane integrity were assessed. The ...

The aim of this study was to evaluate the enzymatic antioxidant capacity of cryopreserved bovine semen with different sources of low-density lipoproteins (LDL) and trehalose (T). Ten ejaculates selected from five bulls were cryopreserved under the treatments: LDL (8% m/v), T (100 mM), TLDL (100 mM T and 8% m/v LDL), centrifuged egg yolk (CEY, 20% v/v) and egg yolk (EY, 20% v/v) as control treatment. After the semen thawing process, the antioxidant enzymatic capacity of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) of bovine semen were evaluated by spectrophotometry plus its relationship with spermatic kinetic parameters. A completely random model was used. The normality of the variables was validated by the Kolmogorov- Smirnov test. The sources of variation were evaluated through a mixed model. The comparison of the adjusted means between the treatments was ...

The development rates and the quality of the bovine embryos produced in vitro are lower than those produced in vivo, due to the oxidative stress to which they are subjected during their manipulation and the culture conditions. The aim of this study was to evaluate the production, cellularity and vitality of bovine blastocysts produced in vitro with different concentrations of quercetin. A total of 2108 oocytes were matured in vitro in medium 199 with 10% FBS, 5.0 μg/ml LH and 0.5 μg/ml FSH. Fertilization was carried out in TALP medium with 2x106 spermatozoa/ml. Potential zygotes were cultured at 38.7 °C with 5% CO2 in SOF culture medium supplemented with six concentrations of quercetin (1, 5, 10, 15, 20, 50 μM) and as controls were used the SOF medium without quercetin and supplemented with DMSO. The rates of cleavage (day 2) and blastocysts (day 7) were determined, as well as the ce...

Se desconoce en gran medida el aporte del plasma seminal a la criotolerancia del semen equino. Algunos componentes del plasma seminal son conocidos por su acción antioxidante y, en consecuencia, podrían influir en la estabilidad oxidativa de los lípidos y las proteínas del semen. Esta investigación tuvo como objetivo evaluar la relación entre la estabilidad oxidativa y algunos componentes del plasma seminal con la criotolerancia del semen equino. En el plasma seminal de 30 caballos se evaluó el contenido de vitamina C (VC) por HPLC, proteínas totales (PT) por Bradford, y la oxidación proteica (OP) y lipídica (PL) por medición de carbonilos y el ensayo TBARS, respectivamente. El semen se congeló con 10% de plasma seminal y se evaluó la calidad seminal posdescongelación. Se realizó el ajuste de modelos lineales generalizados (GLM), análisis de regresión y correlación, y l...

Extenders have been established as an alternative to compensate the antioxidant deficiencies of stallion semen during freezing. Several research works have aimed to evaluate the effect of various additives with antioxidant properties on the equine sperm cryotolerance; however, in limited occasions this has been linked with the extender total antioxidant capacity (TAC). This research aimed to evaluate the effect of two additives on freezing extender TAC and stallion semen cryotolerance. Semen of five Colombian Creole horses was frozen in an extender under three treatments: T0: control, T1: quercetin 100 μM, T2: phosphates and carbonates (Na2HPO4 0.32 mg/ml + KH2PO4 0.67 mg/ml + K2CO3 0.39 mg/ml). TAC of extender was evaluated by the ABTS test. Post-thaw motility and kinetics, abnormal morphology (AM), and structural (MSI) and functional (HOS) plasma membrane integrity were assessed. The ...

The aim of this study was to evaluate the enzymatic antioxidant capacity of cryopreserved bovine semen with different sources of low-density lipoproteins (LDL) and trehalose (T). Ten ejaculates selected from five bulls were cryopreserved under the treatments: LDL (8% m/v), T (100 mM), TLDL (100 mM T and 8% m/v LDL), centrifuged egg yolk (CEY, 20% v/v) and egg yolk (EY, 20% v/v) as control treatment. After the semen thawing process, the antioxidant enzymatic capacity of catalase (CAT), superoxide dismutase (SOD) and glutathione peroxidase (GPx) of bovine semen were evaluated by spectrophotometry plus its relationship with spermatic kinetic parameters. A completely random model was used. The normality of the variables was validated by the Kolmogorov- Smirnov test. The sources of variation were evaluated through a mixed model. The comparison of the adjusted means between the treatments was ...

The development rates and the quality of the bovine embryos produced in vitro are lower than those produced in vivo, due to the oxidative stress to which they are subjected during their manipulation and the culture conditions. The aim of this study was to evaluate the production, cellularity and vitality of bovine blastocysts produced in vitro with different concentrations of quercetin. A total of 2108 oocytes were matured in vitro in medium 199 with 10% FBS, 5.0 μg/ml LH and 0.5 μg/ml FSH. Fertilization was carried out in TALP medium with 2x106 spermatozoa/ml. Potential zygotes were cultured at 38.7 °C with 5% CO2 in SOF culture medium supplemented with six concentrations of quercetin (1, 5, 10, 15, 20, 50 μM) and as controls were used the SOF medium without quercetin and supplemented with DMSO. The rates of cleavage (day 2) and blastocysts (day 7) were determined, as well as the ce...

The reproductive activity of various species of mammals is influenced by the sperm subpopulations of their ejaculates. These subpopulations differ in the motility and kinetic characteristics of the sperm and have been related and classified according to their capacity for the fertilization process. The objective of this research was to evaluate the effect of the composition of the semen diluent on the sperm subpopulations of the chilled rabbit semen. The semen of 10 rabbits of the Californian breed (three ejaculated per animal) was obtained. Each ejaculate was divided into four aliquots, which were diluted in one of four diluents, whose basic composition consisted of T1: skim milk and sugars; T2: dextrose, sodium citrate and potassium acetate; T3: sodium caseinates, phosphates and sugars; and T4: tris-citric acid and egg yolk. Each aliquot was kept refrigerated at 16 °C for 72 h. I...

The aim of this study was to evaluate the post-thawing quality of equine semen subjected to two dimethylformamide (DMF) addition schemes during freezing. Semen of five Colombian Creole horses was obtained and diluted in modified INRA82® medium with two DMF addition treatments, T1: immediate addition of 5% DMF, and T2: fractional addition of 5% DMF (during 10 min). Semen was packed into 05. ml straws and subjected to conventional freezing. Sperm motility, vitality and morphology before and after dilution were assessed. In addition, a post-thaw analysis of sperm motility and kinetics using a computerized system and the evaluation of plasma membrane integrity by the hypoosmotic test (HOS) were performed. Before and after freezing, a lower reduction in sperm motility was observed by the immediate addition of DMF (T1), compared to the fractional addition of the cryoprotectant (T2) (p<0.05...