[Effect of insulin and leptin on ram sperm capacitation for in vitro embryo production]

Improvement of sperm quality as a research field in reproductive biotechnology of domestic animal can be considered as a key element for in vitro fertilization. The aim of the present study was to investigate the effects of insulin and leptin on ovine sperm capacitation / acrosomal reaction, viability and fertilization. The semen samples of 10 Bakhtiari rams were collected by artificial vagina. Using dose response study, the most efficient doses of insulin and leptin were chosen. Each sample was assigned to four experimental groups including insulin [1nM], leptin [100nM], mixed of leptin-insulin and control [without hormone]. Sperm capacitation/acrosomal reaction, viability and fertilization were evaluated by chlortetracycline staining, eosin-negrosin and in-vitro fertilization methods, respectively. values were compared among groups by 1-way ANOVA. Values of capacitation/ acrosomal reaction rate showed significant increase in response to insulin and leptin at 30, 60 and 120 min time points. The sperm viability was significantly [p<0.05] increased in response to insulin when compared with the control group at 30 min time point, without any effect in the other time points. On the other hand, insulin and leptin did not show significant effect [p>0.05] on sperm fertilization. This study indicated that insulin and leptin improved ram sperm capacitation / acrosomal reaction and viability while their effects on in vitro embryo production were inconsiderable

Seroprevalence of Toxoplasma gondii infection in dogs in Tehran, Iran

Toxoplasma gondii infects a wide range of animals; felines are definitive hosts and other animals including the dogs are intermediate hosts. The aim of this study was to determine the seroprevalence of T. gondii infection in dogs in Tehran, capital of Iran and to investigate possible associated risk factors. Three hundreds ninety six serum samples were collected during 2007-8 from the dogs. Collected samples were tested using an indirect fluorescent antibody test [IFAT] in dilutions of 1:16 and more. All procedures were carried out in Shahrekord University, Iran. All the data were analyzed using SPSS software, qui square test with confidence interval of 0.95. From evaluated samples, 89 [22.47%] were positive in titers of at least 1:16. further evaluations in other dilutions showed positive results in dilutions of maximum 1:16, 1:32, 1:64, 1:128 and 1:256 in 38, 29, 15, 2 and 5 dogs respectively. Investigation of the role of risk factors showed no sex predisposition while infection rate was significantly higher in dogs older than one year old. Living places were of significant importance; infection rate was significantly higher in stray or guard dogs in compare with household dogs [P<0.05]. Relatively high seroprevalence of T. gondii infection in dogs in Tehran shows high environmental contamination. It is recommended that the dogs with suspected clinical signs be tested for T. gondii infection

In vitro maturation of dromedary camel oocytes in Hamsvs F10 medium supplemented with different concentrations of fetal bovine serum

To mature dromedary camel oocytes for using them in an IVF system. Design: Interventional study. Ovaries from dromedary camels in local slaughterhouses. Removing varies from camels in a local slaughterhouse, carrying them to the laboratory in warm saline solution, aspiration of follicles, isolation and transferring of oocytes into TCM-199 and Ham's F10 supplemented with 0-10% heat inactivated fetal bovine serum [FBS], culturing oocytes for up to 24h in a COz incubator. After culture oocytes were denuded and put into PBS containing 0.1% hyaluronidase and passing through a fine pipette. Oocytes were then mounted onto slide glass and fixed and stained for evidence of maturation. ANOVA and when a significance different was seen, Duncan's Multiple Range Test. When oocytes from fresh ovaries were culture in Ham's F10 without protein, only 17.65% of them reached to MIL However, significantly [P<0.05] higher oocytes reached to Mil in 5 and 10% PCS [36.84% and 33.33% for 5 and 10% FBS respectively], which were not dose dependent. When cool stored ovaries were used for oocyte maturation, 14.54% of oocytes reached to MIL In protein-free medium However, significantly [P<0.05] higher oocytes reached to Mil in 5 and 10% PCS [25.86% and 33.33% for 5 and 10% FBS respectively]. Although increasing the protein increased the maturation rates, the difference was not significant. Under the present condition it seems that cool stored ovaries could be used for in vitro maturation of camel oocytes

Effects of different concentrations of aflatoxin B on ram epididymal and ejaculatory sperm viability and motility in vitro

To study the effects of aflatoxin on ram epididymal and ejaculatory sperm cells. Interventional study. 10 Chall rams and 25 isolated testicles. Chall ram testicles [n=25] were obtained from slaughter-house, cauda epididymides were incised, sperm samples were isolated and put into media with increasing concentrations of Aflatoxin B. Ejaculates were obtained from 10 healthy Chall rams and the same procedure was assigned. Every hour sperm cells were objected to live-dead staining using eosin - nigrosin procedure and examined under an optic microscope at magnification of xl00, Motility was also assessed in the same time using warm slide glass and magnification of x 10-40. Statistical analysis: ANOVA and Duncan's multiple range test. While after one hour incubation viability of ejaculatory and epididymal sperm cells were 81.25 and 83.24%, when aflatoxin was added [7.81, 31.25 and 62.6 ppb] these values drastically reduced back [p<0.05] in a concentration dependent manner for both epididymal [72.92, 71.8 and 66.72%] and ejaculatory [72.48, 69.6 and 63.63%] sperm cells. During 5 h incubation, viability decreased moderately in all groups. However differences among groups remained unchanged. Furthermore, epididymal sperm motility in the 1st h incubation was significantly higher [p<0.05] than those values in treatment with of 31.25 and 62.6 ppb aflatoxin [51.87 and 15.93%]. Ejaculatory sperm motility was 93.98% control group [93.98%] was significantly higher than those values in treatment with of 31.25 [52.09%] and 62.6 [18.09%] ppb aflatoxin. In spite of differences among groups, values were more apparent for epididymal sperm. Aflatoxin has detrimental effects on sperm viability and motility. However, its effect on motility is more severe