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Articles by H. Wahid
Total Records ( 7 ) for H. Wahid
  Hajarian Hadi , H. Wahid , O. Abas Mazni , Y. Rosnina , M. Daliri , M. Dashtizad , A. Faizah , K. C. Yap , F. J. Fahrul and A. Fazly
  Problem statement: Vitrification is replacing conventional slow freezing to cryopreserve gametes and embryos especially for in vitro production of embryo in domestic animal species. However, the results are still not satisfactory. The aim of this experiment was to study the effect of different equilibration temperatures on in vitro viability of immature bovine oocytes after vitrification. Approach: Oocytes were obtained from slaughterhouse ovaries. Only grade one oocytes were used. Oocytes were equilibrated in three different temperatures: 32, 37, or 41°C. Immature oocytes were equilibrated in VS1 (7.5 Ethylene Glycol (EG) + 7.5% DMSO) for 10-12 min and then exposed to VS2 (15% EG + 15%DMSO + 0.5M sucrose) for one min. Thereafter oocytes were loaded on hand-made Cryotop and directly plunged into liquid nitrogen. After warming, oocytes were examined for viability, maturation, cleavage and blastocyst production. Results: Oocytes that were equilibrated at 37°C had significantly higher (p<0.05) viability than 41°C, but there were no significant difference between 37 and 41 with 32°C. Maturation rate in 37°C group was significantly higher compared with other groups. The highest percentage of degenerated and germinal vesicle stage oocytes were obtained from 41°C than 32 and 37°C. Cleavage rate of 37°C group (38.77%) was greater than other groups (30.84 and 28.95% for 32 and 41°C, respectively). The highest blastocyst rate was also produced when oocytes equilibrated at 37°C (6.45%). Conclusion: In conclusion, these results indicated that immature bovine oocytes can be equilibrated successfully at 37°C while higher or lower temperature can significantly decrease their subsequent viability and development.
  Hajarian Hadi , H. Wahid , Y. Rosnina , M. Daliri , M. Dashtizad , H. Karamishabankareh , A. Faizah , M.I. Iswadi and O. Abas Mazni
  Beside cooling/warming rates and composition of vitrification solution, developmental stage of immature oocytes may also affect their vitrification outcome. The aim of the present study was to evaluate the selection effect of developmentally competent immature bovine oocytes by Brilliant Cresyl Blue (BCB) on maturity of oocytes after vitrification. Oocytes were obtained from slaughterhouse ovaries. Only oocytes with 4-5 layers of cumulus cells and homogenous cytoplasm were used. After exposure to BCB stain, immature oocytes were divided into colored (BCB+) and colorless (BCB-) cytoplasm groups. Immature oocytes were equilibrated in VS1 (7.5 Ethylene Glycol (EG)+7.5% DMSO) for 10-12 min and then exposed to VS2 (15% EG+ 15% DMSO+0.5M sucrose) for 1 min. Thereafter, oocytes were loaded on Cryotop and directly plunged into liquid nitrogen. After warming, oocytes were examined for presence of polar body and nuclear maturity. Higher number of oocytes in BCB+group extruded first polar body in comparison with other vitrified groups but not significantly (p>0.05). Compared to the BCB- oocytes, there was significantly lower percentage of degeneration for BCB+oocytes (p<0.05). Within vitrified groups, reaching to the MII stage was significantly higher in BCB+group (51.5%) compared with BCB and vitrified-control groups (27.9 and 40.3%, respectively). These results indicated that selection of potent immature bovine oocytes using brilliant cresyl blue improved the nuclear maturity of immature oocytes after vitrification. In addition, this selection can be a valuable tool to improve the vitrification outcome.
  H. Hajarian , H. Wahid , Y. Rosnina , M. Daliri , M. Dashtizad , T. Mirzapour , N. Yimer , M.M. Bukar , M.I. Iswadi and O. Abas Mazni
  The aim of this study was to evaluate the effectiveness of different cryodevices (Open Pulled Straw (OPS), Electron Microscopy Grid (EMG) and cryotop for vitrification of immature bovine oocytes. Polar body, MII stage, survivability and subsequent developmental rates were compared. Only oocytes with 4-5 layers of cumulus cells were used. Oocytes were equilibrated in the first vitrification solution (VS1; HS+10% DMSO+10% Ethylene Glycol (EG)) for 30-45 sec and then in the second vitrification solution (VS2; 20% DMSO+20% EG+0.5 M Sucrose) for 25 sec. Within 30 sec they were mounted on one of the cryodevices and directly plunged into Liquid Nitrogen (LN2) for 10 days. Immature oocytes vitrified using cryotop represented higher rate of polar body extrusion and nuclear maturity (p<0.05). The highest survivability resulted from cryotop and EMG groups and no significant difference found between them. Vitrified oocytes in cryotop group had highest cleavage and blastocyst rates. All of the mean measured rates for vitrified/warmed immature oocytes were significantly lower than that of control group (p<0.05). In conclusion, results of this study showed the superiority of cryotop device for vitrification of immature bovine oocytes which resulted in higher viability and subsequent embryo development.
  M. Dashtizad , H. Wahid , O. Abas Mazni , Y. Rosnina , M. Daliri and H. Hajarian
  Development of efficient culture system to support embryonic development would be valuable when percentage of produced embryos reaching to the blastocyst stage is important. However, the rate of bovine embryo production in vitro is still lower than expected. Present study was performed to investigate the effect of ghrelin on nuclear maturation and subsequent bovine embryo development in vitro. Cumulus-oocyte-complexes were collected from slaughterhouse ovaries and randomly allocated in each treatment groups. Five different concentrations of ghrelin (0, 5, 50, 500 and 1000 ng mL-1) were added to the in vitro maturation medium (Hepes-buffered medium 199+fetal calf serum+gonadotrophins+insulin+antibiotics). The proportion of oocytes developed to metaphase II stage was significantly increased at 5 and 50 ng mL-1 ghrelin (86.32±3.38 and 89.77±2.92%, respectively). The result also indicated that adding high concentration of ghrelin adversely affect (p<0.05) the nuclear maturation rates of bovine oocytes. However, the subsequent embryo development was not significantly affected by addition of ghrelin to the IVM medium. This study showed that inclusion of 5-50 ng mL-1 ghrelin in maturation medium may have beneficial effects on nuclear maturation of bovine oocytes in vitro.
  N.P. Khanh , Y. Rosnina , M.A. Omar , G.K. Dhaliwal , H. Wahid , A.M. Khumran , K.C. Yap , M. Fahmi and A. Azmil
  The objective of this experiment is to describe the influences of biostimulation on oestrus behaviour, ovulation time, progesterone levels and conception rate in beef cows. Three groups of cattle were investigated; CNB, PB and MB which comprised 17 multiparous, 12 primiparous and 13 multiparous cows, respectively. On the 3rd day of CIDR insert, three bulls were introduced into the MB and PB groups for biostimulation and breeding. CNB cows were AI 12 h after standing heat. Blood samples were collected every 3 days from CIDR insertion to end of experiment. Ovulation time and pregnancy were determined by ultrasound twice daily for 4 consecutive days and on day 35 after CIDR removal, respectively. Overall, mean percent of cows in oestrus was 90.52%. PB and MB scored higher than CNB for oestrus intensity. Mounting another cow being mounted but not standing and standing oestrus were commonly exhibited during oestrus. Oestrus duration was significantly longest in PB. Conception rate in PB and MB were three times higher than CNB. Only primiparous cows displayed longer, more intense oestrus and longer ovulation time. Conception rate is higher by natural mating than AI.
  A. Azizah , O.M. Ariff , H. Yaakub , J. Ahmad , S. Sukardi and H. Wahid
  The study was conducted to determine the differences in the interval between removal of Controlled Internal Drug Release (CIDR) and standing oestrus (C_ES) interval between CIDR removal and Ovulation (C_OV) interval between oestrus and Ovulation (ES_OV), predetermined Artificial Insemination (AI) and Pregnancy rate following prostaglandin (PGF) treatment and CIDR removal at standing oestrus in Kedah-Kelantan (KK) and KK crossbred cows. A total of 35 KK (n = 11), Brakmas (BK: n = 10) and Charoke (CK: n = 14) cows were inserted with CIDR containing 1.38 g Progesterone (P4) for 7 days and followed with intramuscular injection of 500 μg Cloprosterol of PGF synthetic analogue 2 days prior CIDR removal. The oestrous behaviour was observed for 72 h at 2 h intervals beginning 12 h after CIDR removal. AI was carried out at 6 and 12 h after standing oestrus was detected. Mean interval between C_ES, C_OV and ES_OV were 52.97±6.16, 94.42±16.41 and 53.75±25.36 h, respectively. The number of follicles >5 mm in diameter at the time of CIDR removal was significantly lower in CK (p = 0.04) compared to KK. The results of POF size and number of follicle size ≥5 mm either at CIDR removal or prior ovulation and P4 concentration at day 0, 7 and 14 were found to be not significantly different at p = 0.05. The pregnancy rate was higher (p<0.05) in CK (43.8%) followed by KK (31.3%) and BK (25%). The study provides results which could be used in the development of TAI on oestrous synchronisation protocol of KK and KK crossbred cows.
  A.M. Khumran , Y. Rosnina , M.O. Ariff , H. Wahid , G. Dhaliwala , N.P. Khanh , K.C. Yap , M. Fahmi and M.E. Azmil
  The aim of this study was to compare the estrus response and pregnancy rate between the indigenous beef cows of Malaysia; Kedah-Kelantan (KK) and the exotic beef cows; Brangus (BR) following progesterone and prostaglandin-based estrus synchronization treatments. A total of 40 KK and 30 BR open cows were selected and each breed group was randomly divided equally into two. Cows in KK1and BR1 were treated with estradiol benzoate (Cidirol, 1 mg, im) each at the time CIDR® was inserted (Day 0). Cloprostenol (Estrumate, 250 μg, im) was administered at the time of CIDR® removal on Day 9 while 1 mg of Estradiol Benzoate (EB) was injected on Day 10. On the other hand, KK2 and BR2 cows received intramuscular injections of 500 and 250 μg of cloprostenol, on Day 0 and 11, respectively. All cows were then observed for estrus signs and scanned per rectum for ovulation followed by AI upon detection of estrus. Pregnancy status was diagnosed 45 days after AI. Both treatments were effective in inducing observable estrus in all groups with synchrony of ovulation resulting in CL development and pregnancy. In the progesterone-based treatment groups, 84.2% of KK1 and 78.8% of BR1 responded. In the prostaglandin-based treatment groups, KK2 responded with the highest proportion (80.0%) compared with BR2 (50.0%). However, there was no significant difference in rate of ovulation (84.2 vs. 64.3%; 70.0 vs. 42.9%) and pregnancy (31.6 vs. 14.3%; 45.0 vs. 21.4%) among all the four experimental groups. The interval to ovulation from the last treatment time varied significantly among all the treatment groups with a higher variation observed in BR, ranging from 48 h when treated with CIDR to 84 h after treatment with PGF. These variations could be explained by the difference in ovarian status at the time of treatment. In conclusion, the result of this data showed KK cows had a better rate of ovulation and pregnancy than BR cows in both treatments though not statistically significant. It can therefore be gathered that KK and BR responded effectively to estrus synchronization and produce acceptable pregnancy rates by both progesterone and prostaglandin-based protocols for breeding and genetic improvement.
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