Although, it has been success in the generation of animal clones from somatic cells in various animal species, the information related to nuclear reprogramming of cloned embryos is found to be limited. This study aims to compares the effect of both Scriptaid (SCR) and Trichostatin (A) treatments in improving cloning efficiency, and embryos developmental rate of cloned sheep embryos in vitro. Three groups were formed, i.e., one SCR group, second TSA group, with both treatment concentrations of 5 nM, 50 nM, and 500 nM, respectively, and third were control group with 0 nM. Methods: Ovaries of slaughtered sheep were collected and oocytes were recovered from antral follicles using aspiration method and in vitro maturation of oocytes were done. Then zona dissecting with micropipettes and oocyte enucleation were carried out under the micromanipulator. Later nuclear transfer, cell fusion and activation were done via cell fusion machine. Finally the embryo cultured in incubating chamber at the CO2 incubator up to 9 days. The result: In general the results showed that when the concentration increases the cleavage rate increased. The cleavage rates of the SCNT embryos treated with SCR at different concentrations are closely related to cleavage rate of embryos treated with TSA at same concentration; such as 39.47% for 500 nM TSA, 38.09% for 500 nM SCR; 18.6% for 50 nM TSA, 19.17% for 50 nM SCR, and 22.64% for 5 nM TSA, 17.18% for 5 nM SCR. As for the control group, the cleavage rate of the SCNT embryos cleavage ratewere27.47%., 30% and 30.85% respectively for bothtreatments. While there is a significant difference in TSA treatments at an eight-cell stage at the concentration (5 and 50 nM TSA) compared to the all other cleavage cell stages of (500 nM TSA and control). Also their were a differences between (50 nM of TSA) compared to the (50 nM SCR). Also there were a significant differences between the 16 cell stage at the (500 nM TSA) compared to other treatment (5 nM, 50 nM TSA and control). Regarding the SCR there were a significant difference at 8 cell stage between (5 nM SCR), compared to the other treatment (50 nM, 500 nM SCR and control). Also there were a significant difference at 16 cell stage between (50 nM, and 500 nM SCR), compared to the other treatment (5 nM SCR and control). While in the development of the embryos reach to blastocyst stage the SCR and the control group show a higher rate, in compered to TSA that did not show any development to blastocyst stage. The total SCR treatment showed (3/41 = 7.31%), and the total control showed (4/89 = 4.49%) blastula stage. It concludes that SCR improve the final development blastula stage compared to the TSA treatments that did not improved embryos reach to final developmental blastula stages may be due to spices differences or to the toxicity of TSA, especially at higher concentrations.

The aim of this study is to evaluate the effect of sheep follicular fluid (SFF) supplementation of the in vitro maturation (IVM) media of sheep oocytes on the resumption of meiosis, glutathione (GSH) level, and expression of apoptosis (Bax, Bcl-2) as well as heat shock protein beta-1 (HSPB1) genes. Sheep ovaries were collected from the central slaughterhouse of Riyadh city, KSA. Oocytes were aspirated from 3 to 8 mm follicles. Sheep oocytes were cultured in maturation medium with different concentrations of sheep follicular fluid: 0% (control), 10%, 20% and 40% for 24 h. The results indicated that the maturation rate of oocytes was significantly (p ≤ .05) decreased in 40% SFF (36.87%) versus the control (61.3%), 10% SFF (63.95%) and 20% SFF (64.08%). The supplementation of the IVM medium with 10% SFF induced an intra-oocyte GSH concentration that was significantly higher than in sheep oocytes cultured with 20% and 40% SFF and similar to the GSH content in oocytes cultured without SFF. Real-time polymerase chain reaction analysis of gene expression revealed no significant differences in the Bax and HSPB1 genes between the control and 10% SFF, whereas they were significantly higher in 40% FF (p ≤ .05) compared to the control. The expression of Bax:Bcl-2 was significantly higher in 20% and 40% SFF compared to the control group. In conclusion, the addition of SFF to the IVM culture of sheep oocytes is recommended to support nuclear maturation and increase oocyte competence.

Background: Embryo production in vitro requires three steps: in vitro maturation (IVM), in vitro fertilization (IVF) and in vitro culture (IVC). The first step in the in vitro maturation of oocytes is germinal vesicle breakdown (GVBD), followed by completion of the 1st meiotic division and formation of the 1st polar body. These parameters are critical during ovary and oocyte selection. This study aimed to evaluate bovine ovary and oocyte collection for IVM. Methods: Ovaries collected from cows in a 0.9% NaCl saline solution at the central slaughterhouse were divided into two normal and abnormal ovaries according to their morphological appearance. The morphometric dimensions of bovine ovaries, such as weight and deamination (length, width and volume), were recorded. The oocyte was extruded and its deamination occurred before and after culture in maturation medium. Result: The mean ovary weight, volume, length and width were not significantly different between the two ovary types. Additionally, the dominant and subordinate follicle diameters in both ovary types showed no significant differences. Furthermore, the oocyte number per ovary of the normal and abnormal oocytes showed no significant differences. The mean cumulus oocyte complex before maturation showed no significant difference (52.73±7.23 mm for the normal ovary vs. 43.015±5.41 mm for the abnormal ovary). However, after maturation, a highly significant difference was found (P less than 0.001) between the normal ovaries before and after maturation (178.10±15.36 µm) and abnormal ovaries (10.45±7.99 µm). Additionally, data analysis of oocytes with or without 1st polar bodies revealed a highly significant difference (P less than 0.001) between oocytes of the normal ovary with (32.15±4.19) and without (10.95±1.59) 1st polar bodies and oocytes of the abnormal ovary with (7.0±078) and without 1st polar bodies (3.3±0.32). Thus, the critical point at which the normal ovary produces better in vitro matured follicles with good oocyte quality and produces the 1st polar body determines developmental competence. Therefore, the best selection of normal ovaries will enhance in vitro maturation for subsequent experiments, such as in vitro fertilization or cloning and in vitro embryo development.

The immune and antioxidant genetic factors that could converse with mastitis susceptibility in dromedary camels were looked at in this research. Of 120 female dromedary camels (60 healthy, and 60 with mastitis) were utilised. Each camel’s jugular vein was pierced to obtain five millilitres of blood. The blood was placed within tubes containing sodium fluoride or EDTA anticoagulants to obtain whole blood and extract DNA and RNA. The immunological (OTUD3, TLR2, TLR4, STAB2, MBL2, TRAPPC9, and C4A) and antioxidant (CAT, SOD3, PRDX6, OXSR1, NDUFS6, SERP2, and ST1P1) genes’ nucleotide sequence polymorphisms between healthy and mastitis affected she-camels were discovered using PCR-DNA sequencing. Fisher’s exact test revealed that camel groups with and without mastitis had noticeably different odds of all major nucleotide alterations propagating (p < 0.01). Mastitic camels were significantly more likely to express the OTUD3, TLR2, TLR4, STAB2, MBL2, TRAPPC9, C4A, OXSR1, SERP2, and ST1P1 genes (p < 0.05). However, CAT, SOD3, PRDX6, and NDUFS6 genes elicited a different pattern. The results may be used to develop management strategies and support the significance of nucleotide differences and gene expression patterns in these markers as indicators of the incidence of mastitis.

In an innovative approach to push the boundaries of antimicrobial and antioxidant strategies, we present the synthesis and characterization of a novel terpolymer derived from N-Phenyl-p-phenylenediamine and 2-aminopyrimidine with formaldehyde in the presence of dimethylformamide as a reaction medium through polycondensation technique. Leveraging this terpolymer as a ligand, we introduce an intriguing terpolymer-metal complex, created with Ni (II) metal ion. In our pursuit to validate the structure and properties of these substances, we performed meticulous characterizations using important spectral studies such as FTIR, electronic, and ¹H NMR spectroscopy. This provided us with a unique fingerprint for the (N-Phenyl-p-phenylenediamine-2-aminopyrimidine-formaldehyde) terpolymeric ligand (PAF) and its metal complex. In addition, the molecular weights of PAF terpolymer were established using gel permeation chromatography.