Aim:To monitor the effect of nutrition and pregnancy on oxidative status of animals under the arid condition of South Sinai.Materials and Methods:Blood samples were taken from two groups of animals: The first group retained in farm and fed on concentrate (high diet) and another group grazing natural forage (low diet). Each group was subdivided into pregnant and non-pregnant animals. Blood samples were assayed for their content of malondialdehyde (MDA), total antioxidant capacity (TAC), catalase (CAT), and superoxide dismutase (SOD) enzymes.Results:MDA level significantly increased in pregnant animals fed either concentrate or grazing low-quality forage and accompanied by a low level of TAC in pregnant grazing animals fed low-quality forage. The activity of CAT decreased in pregnant fed either concentrate or grazing and SOD significant decrease in the pregnant grazing group. These data suggested that the animals might have experienced some degree of oxidative stress and lipid peroxidation and indicating that redox homeostasis was impaired in those pregnant and specially fed on forage rations.Conclusion:Pregnancy constituted the most oxidative stress facing the grazing and concentrated diet feed sheep and goats under arid and saline conditions of Southern Sinai, Egypt.
This study was designed to evaluate effects of different combinations of cryoprotectants on the ability of vitrified immature buffalo oocytes to undergo in vitro maturation. Straw and open-pulled straw (OPS) methods for vitrification of oocytes at the germinal vesicle stage also were compared. The immature oocytes were harvested from ovaries of slaughtered animals and were divided into three groups: (i) untreated (control); (ii) exposed to cryoprotectant agents (CPAs); or (iii) cryopreserved by straw and OPS vitrification methods. The vitrification solution (VS) consisted of 6 m ethylene glycol (EG) as the standard, control vitrification treatment, and this was compared with 3 m EG + 3 m dimethyl sulfoxide (DMSO), 3 m EG + 3 m glycerol, and 3 m DMSO + 3 m glycerol. Cryoprotectants were added in two steps, with the first step concentration half that of the second (and final) step concentration. After warming, oocyte samples were matured by standard methods and then fixed and stained for nuclear evaluation. Rates of MII oocytes exposed to CPAs without vitrification were lower (54.3 +/- 1.9% in EG, 47.5 +/- 3.4% in EG + DMSO, 36.8 +/- 1.2% in EG + glycerol and 29.9 +/- 1.0% in DMSO + glycerol; p < 0.05) than for the control group (79.8 +/- 1.3%). For all treatments in each vitrification experiment, results were nearly identical for straws and OPS, so all results presented are the average of these two containers. The percentages of oocytes reaching telophase-I or metaphase-II stages were lower in oocytes cryopreserved using all treatments when compared with control. However, among the vitrified oocytes, the highest maturation rate was seen in oocytes vitrified in EG + DMSO (41.5 +/- 0.6%). Oocytes cryopreserved in all groups with glycerol had an overall low maturation rate 19.0 +/- 0.6% for EG + glycerol and 17.0 +/- 1.1% for DMSO + glycerol. We conclude that the function of oocytes was severely affected by both vitrification and exposure to cryoprotectants without vitrification; the best combination of cryoprotectants was EG + DMSO for vitrification of immature buffalo oocytes using either straw or OPS methods.
Bone morphogenetic protein 15 (BMP15/FecX) gene is considered one of the major genes and a candidate marker for the reproduction in farm animals, especially sheep. The present study aimed to detect the genetic polymorphisms of BMP15 gene in sheep using PCR-RFLP technique. In the present study, 115 ewes were assigned into high and low prolificacy categories according to their reproductive history. In high prolific group (n = 20), ewes produced twins more than single births. In the low prolific type (n = 95), the ewes produced single births more than twins. DNA was extracted from blood samples of all ewes, subjected to PCR-RFLP analysis and confirmed by sequence analysis. The PCR products of 356 bp size were cut with HinƒI restriction enzyme. Three digested fragments of 70, 117 and 169 bp were obtained in both types of sheep. All animals were homozygous with CC genotype. In conclusion, the accessible findings did not detect any mutation in FecX gene in sheep, regardless their prolificacy. Therefore, further attempts are necessary to detect other SNP for BMP-15 gene in Egyptian sheep breeds.
HIS WORK aimed to search for single nucleotide polymorphisms of gonadotropin releasing hormone receptor (GnRHR) gene in Egyptian buffalo which involved in endocrine control of fertility. An ultrasonographic examinations of anestrum and repeat breeders' buffaloes were performed prior to blood sampling through a transrectal ultrasonography. DNA was extracted from blood and the PCR and singlestrand conformation polymorphism (SSCP) techniques were adopted to study of GnRHR gene polymorphism. The PCR amplified a fragment with 240-bp in size and the SSCP results showed that there is a genetic polymorphism with three different patterns (AA, BB and CC) in Egyptian buffalo. The CC genotype was associated with smooth inactive ovary, while BB genotype was associated with fertility in buffalo. Multiple sequence alignment of the three patterns sequences revealed that GnRHR had 5 single nucleotide polymorphisms including one nucleotide insertion, one nucleotide deletion and 3 nucleotide substitutions. Insertion was at position 4 in pattern III. The deletion was at position 189 in pattern III, while the 3 nucleotide substitutions were at positions 204 (T/G), 206 (T/A) and 207 (A/T). In conclusion, GnRHR gene could be used as a candidate marker for fertility in Egyptian buffaloes with its mutation is related to ovarian inactivity.
Fresh semen from fifty bulls was evaluated for ejaculate volume, individual motility, concentration, lives sperm and abnormalities as well as acrosome integrity. Bulls were classified according to semen motility into two groups; good and poor. DNA was extracted from semen of both groups, then the PCR followed by single-strand conformation polymorphism (SSCP) techniques were performed for mutation detection in gonadotropin releasing hormone receptor (GnRHR) gene through multiple sequence alignment. The results showed that the percentages of sperm motility, sperm concentration and live sperm, were significantly higher in good than poor semen quality bulls. However, semen volume, percentage of acrosome integrity and abnormalities did not differ between good and poor. The PCR amplification of 240-bp fragment and the results of SSCP appeared a genetic polymorphism with two patterns. Pattern I was seen in all good bulls, with incidence of 100%. Pattern II appeared only in poor semen quality bulls with a frequency of 31.25%. The sequence analysis of the PCR product for the two patterns showed two single nucleotide polymorphisms (SNPs) as a transversion base substitution mutation at positions 20 (T/A) and 193 (A/T). The GnRHR gene could be used as a genetic marker related to semen quality in buffalo due to the good semen bulls had a unique pattern.
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