Resumo
The aim of this study was to evaluate the effect of in vitro treatment of ejaculated ram spermatozoa with different concentrations of L-arginine at various incubation times on motility, hyperactivity (HA) and acrosome reaction. Freshly ejaculated spermatozoa collected from three rams were pooled and subjected to the swim up technique in modified sperm Tyrode's albumin lactate pyruvate (S-TALP) medium supplemented with different concentrations of Larginine (0.01, 0.02, 0.03,0.04 and 0.05 mM) at 30, 60,90 and 120 min of incubation. Following sperm incubation, the following parameters were examined: motility, hyperactivity (HA) and acrosome reaction (AR). The results showed that irrespective of the concentration, incubation of ram spermatozoa with Larginine for 30min did not significantly affect the motility. However, increase the time of incubation for more than 30 min significantly decreased (P < 0.05) the motility of the spermatozoa as compared to the control. The lowest motility was recorded when spermatozoa were incubated with 0.05 mM L-arginine for 120 min. Treatment of ram spermatozoa with 0.05 mM Larginine resulted in a significant (P < 0.05) increase in HA% immediately after dilution compared to the control. A significant (P < 0.05) increase in total AR% was concomitant to the increase in the concentration of L-arginine with highest AR achieved at 0.04 mM and 90 min incubation. However, increasing the time of incubation to 120 min significantly decreased (P < 0.05) the percentage of spermatozoa with total AR compared to the other incubation times at 0.02, 0.04, and 0.05 mM L-arginine. In conclusion, under our experimental conditions treatment of ejaculated ram spermatozoa with 0.04 mM of L-arginine for 90 min was considered the best concentration of L-arginine to be used for in vitro induction of acrosome reaction.
Assuntos
Masculino , Animais , Arginina/efeitos adversos , Ejaculação , Ejaculação/genética , Ovinos/embriologiaResumo
The aim of this study was to evaluate the effect of in vitro treatment of ejaculated ram spermatozoa with different concentrations of L-arginine at various incubation times on motility, hyperactivity (HA) and acrosome reaction. Freshly ejaculated spermatozoa collected from three rams were pooled and subjected to the swim up technique in modified sperm Tyrode's albumin lactate pyruvate (S-TALP) medium supplemented with different concentrations of Larginine (0.01, 0.02, 0.03,0.04 and 0.05 mM) at 30, 60,90 and 120 min of incubation. Following sperm incubation, the following parameters were examined: motility, hyperactivity (HA) and acrosome reaction (AR). The results showed that irrespective of the concentration, incubation of ram spermatozoa with Larginine for 30min did not significantly affect the motility. However, increase the time of incubation for more than 30 min significantly decreased (P < 0.05) the motility of the spermatozoa as compared to the control. The lowest motility was recorded when spermatozoa were incubated with 0.05 mM L-arginine for 120 min. Treatment of ram spermatozoa with 0.05 mM Larginine resulted in a significant (P < 0.05) increase in HA% immediately after dilution compared to the control. A significant (P < 0.05) increase in total AR% was concomitant to the increase in the concentration of L-arginine with highest AR achieved at 0.04 mM and 90 min incubation. However, increasing the time of incubation to 120 min significantly decreased (P < 0.05) the percentage of spermatozoa with total AR compared to the other incubation times at 0.02, 0.04, and 0.05 mM L-arginine. In conclusion, under our experimental conditions treatment of ejaculated ram spermatozoa with 0.04 mM of L-arginine for 90 min was considered the best concentration of L-arginine to be used for in vitro induction of acrosome reaction.(AU)
Assuntos
Animais , Masculino , Arginina/efeitos adversos , Ejaculação , Ejaculação/genética , Ovinos/embriologiaResumo
Studies were conducted to compare viability of immature buffalo oocytes vitrified in different types of cryoprotectants on the post-thaw morphological appearance, the in vitro maturation and developmental competence of buffalo oocytes. The vitrification solution (VS) consisted of Dulbeccos phosphate buffered saline (DPBS) supplemented with 0.5 M sucrose, 0.4% bovine serum albumin (BSA) and different types of molar (M) concentrations of the cryoprotectants which were composed of either glycerol (G), ethylene glycol (EG) or dimethyl sulfoxide (DMSO) in order to determine the best type of vitrification cryoprotectants. The concentrations tested were 4 M, 7 M and 7M concentration of G, EG and DMSO, respectively. Cumulus oocyte complexes (COCs) were obtained from slaughterhouse ovaries. OocytesAnd their cumulus cells were vitrified immediately after collection .The COCs were pre-equilibrated in 50% of the VS for 3-5 min, then kept in VS for 1 min and loaded in pre-sterilized 0.25 ml straws for 7-10 days of storage in liquid nitrogen. The straws were thawed in warm water at 37°C for 10 s. The COCs were equilibrated in 0.5 M sucrose in modified phosphate buffer (M-PBS) for 5 min and then washed in washing medium (TCM-199 plus 10% FCS). Oocyte-cumulus cells were evaluated for morphological damage. Morphologically normal COCs (Oocyte-cumulus cells) were cultured in vitro and evaluated for maturation. Oocytes were fertilized with frozen-thawed semen capacitated in Brackett and Oliphant (BO) medium containing heparin and caffeine and were evaluated for cleavage and embryonic development. The results revealed that the proportion of buffalo oocytes found to be morphologically normal was significantly (P < 0.05) higher in EG and DMSO than those obtained in G (85.0 and 83.33 vs. 65.0%, respectively). Among the damaged oocytes, cracking of zona pellucida was the most frequent abnormality observed.
Assuntos
Feminino , Animais , Búfalos , Crioprotetores/uso terapêutico , Técnicas de Maturação in Vitro de Oócitos/métodos , Técnicas de Maturação in Vitro de Oócitos/veterinária , Dimetil Sulfóxido , Etilenoglicol , Glicerol , VitrificaçãoResumo
Studies were conducted to compare viability of immature buffalo oocytes vitrified in different types of cryoprotectants on the post-thaw morphological appearance, the in vitro maturation and developmental competence of buffalo oocytes. The vitrification solution (VS) consisted of Dulbeccos phosphate buffered saline (DPBS) supplemented with 0.5 M sucrose, 0.4% bovine serum albumin (BSA) and different types of molar (M) concentrations of the cryoprotectants which were composed of either glycerol (G), ethylene glycol (EG) or dimethyl sulfoxide (DMSO) in order to determine the best type of vitrification cryoprotectants. The concentrations tested were 4 M, 7 M and 7M concentration of G, EG and DMSO, respectively. Cumulus oocyte complexes (COCs) were obtained from slaughterhouse ovaries. OocytesAnd their cumulus cells were vitrified immediately after collection .The COCs were pre-equilibrated in 50% of the VS for 3-5 min, then kept in VS for 1 min and loaded in pre-sterilized 0.25 ml straws for 7-10 days of storage in liquid nitrogen. The straws were thawed in warm water at 37°C for 10 s. The COCs were equilibrated in 0.5 M sucrose in modified phosphate buffer (M-PBS) for 5 min and then washed in washing medium (TCM-199 plus 10% FCS). Oocyte-cumulus cells were evaluated for morphological damage. Morphologically normal COCs (Oocyte-cumulus cells) were cultured in vitro and evaluated for maturation. Oocytes were fertilized with frozen-thawed semen capacitated in Brackett and Oliphant (BO) medium containing heparin and caffeine and were evaluated for cleavage and embryonic development. The results revealed that the proportion of buffalo oocytes found to be morphologically normal was significantly (P < 0.05) higher in EG and DMSO than those obtained in G (85.0 and 83.33 vs. 65.0%, respectively). Among the damaged oocytes, cracking of zona pellucida was the most frequent abnormality observed.(AU)
Assuntos
Animais , Feminino , Búfalos , Crioprotetores/uso terapêutico , Técnicas de Maturação in Vitro de Oócitos/métodos , Técnicas de Maturação in Vitro de Oócitos/veterinária , Vitrificação , Dimetil Sulfóxido , Etilenoglicol , GlicerolResumo
aim of this study was to evaluate the effects of the reproduct ive status of female dromedary camels ( pregnant vs. non - pregnant ) on the chemical composition, hormonal profile and antioxidant capacity of follicular fluid collected from different sized ovarian follicles during the breeding season . One hundred ovaries we re collected at slaughter house from fifty female dromedary camel s . The ovaries were collected in pairs from each animal and allocated into two groups according to the reproductive status of the females ; 25 pairs were obtained from pregnant females and 25 p airs were obtained from non - pregnant animals . The follicles on each ovary were categorized according to their diameter into three categories; small ( 1 - 3 mm), medium (4 - 9 mm) and large ( 10 - 20 mm). Follicular fluid (FF) aspirated from each follicle category from each pair of ovaries w as analyzed . The results showed that the average number of follicles per ovary was greater ( P < 0.05) in the ovaries obtained from non - pregnant females compared to those collected from pregnant ones ( 6.4 ± 1.2 vs . 3.6 ± 0.9, resp ectively). Progesterone concentrations were significantly higher in the follicular fluid collected from all follicle categories in pregnant animals than those obta ined from non - pregnant animals . Glucose concentration s were higher (P < 0.05) in the follicul ar fluid collected from large follicles in the non - pregnant group (64.9 ± 6.1 mg/d l ) than those obtained from the same follicle category in the pregnant ovaries (45.4 ± 4.0 mg/d l ). C oncentrations of malondialdehyde (MDA) were higher (P < 0.05) in the FF co llected from small, medium and large follicles in pregnant ovaries than non - pregnant ones . In conclusion , these data indicate that FF composition differ s according to the reproductive status of the female . In pregnant camels, the p resence of the corpus lut eum on the ovar ies could play an important role not only in the process of follicle growth and development , but also in the concentrations of biochemical metabolites and hormonal profiles in the FF of dromedary camels.
Assuntos
Animais , Folículo Ovariano/anatomia & histologia , Líquido Folicular/citologia , Ovário/anatomia & histologia , Antioxidantes/análise , Camelus/fisiologiaResumo
aim of this study was to evaluate the effects of the reproduct ive status of female dromedary camels ( pregnant vs. non - pregnant ) on the chemical composition, hormonal profile and antioxidant capacity of follicular fluid collected from different sized ovarian follicles during the breeding season . One hundred ovaries we re collected at slaughter house from fifty female dromedary camel s . The ovaries were collected in pairs from each animal and allocated into two groups according to the reproductive status of the females ; 25 pairs were obtained from pregnant females and 25 p airs were obtained from non - pregnant animals . The follicles on each ovary were categorized according to their diameter into three categories; small ( 1 - 3 mm), medium (4 - 9 mm) and large ( 10 - 20 mm). Follicular fluid (FF) aspirated from each follicle category from each pair of ovaries w as analyzed . The results showed that the average number of follicles per ovary was greater ( P < 0.05) in the ovaries obtained from non - pregnant females compared to those collected from pregnant ones ( 6.4 ± 1.2 vs . 3.6 ± 0.9, resp ectively). Progesterone concentrations were significantly higher in the follicular fluid collected from all follicle categories in pregnant animals than those obta ined from non - pregnant animals . Glucose concentration s were higher (P < 0.05) in the follicul ar fluid collected from large follicles in the non - pregnant group (64.9 ± 6.1 mg/d l ) than those obtained from the same follicle category in the pregnant ovaries (45.4 ± 4.0 mg/d l ). C oncentrations of malondialdehyde (MDA) were higher (P < 0.05) in the FF co llected from small, medium and large follicles in pregnant ovaries than non - pregnant ones . In conclusion , these data indicate that FF composition differ s according to the reproductive status of the female . In pregnant camels, the p resence of the corpus lut eum on the ovar ies could play an important role not only in the process of follicle growth and development , but also in the concentrations of biochemical metabolites and hormonal profiles in the FF of dromedary camels.(AU)