Rosnina Yusoff

Two-dimensional polyacrylamide gel electrophoresis of Bali bull (Bos javanicus) seminal plasma proteins and their relationship with semen quality

The present study evaluated the relationship between Bali bull (Bos javanicus) seminal plasma proteins and different semen quality parameters. Semen samples from 10 mature Bali bulls were evaluated for conventional semen parameters (general motility, viability, and normal morphology), sperm functionality (acrosome reaction, sperm penetration rate, sperm penetration index), sperm kinetics (computer-assisted semen analysis parameters such as sperm velocity), and sperm morphology (acrosome and membrane integrity). Frozen-thawed semen with higher sperm motility, viability, acrosome integrity, and membrane integrity (P < 0.05) are consistently higher in acrosome reaction and sperm penetration assay. Three bulls showed the highest, four bulls displayed the medium, and the remaining three bulls showed the lowest for all sperm parameters and SPA. The proteome maps of seminal plasma from high-quality and low-quality Bali bulls were also established. Seminal plasma of both high-quality and low-quality Bali bulls was subjected to two-dimensional SDS-PAGE with isoelectric point ranged from 3 to 10 and molecular weight from 10 to 250 kDa. Approximately 116 spots were detected with Blue Silver stain, and of these spots, 29 were selected and identified by MALDI-TOF/TOF-MS/MS. A majority of the proteins visualized in the seminal plasma two-dimensional maps was successfully identified. An essential group of the identified spots represented binder of sperm 1 (BSP1), clusterin, spermadhesin, tissue inhibitor of metalloproteinases 2 (TIMP-2), and phospholipase A2 (PLA2). Other proteins found in high abundance included seminal ribonuclease, serum albumin, cationic trypsin, and peptide similar to β2 microglobulin. Thus, a reference map of Bali bull seminal plasma proteins has been generated for the very first time and can be used to relate protein pattern changes to physiopathologic events that may influence Bali bull reproductive performance.

Effect of docosahexanoic acid on quality of frozen-thawed bull semen in BioXcell extender

This study was conducted to investigate the effect of docosahexanoic acid (DHA) supplementation in BioXcell extender on the quality of frozen-thawed bull semen. Twenty-four ejaculates were collected from three bulls (eight from each bull). Ejaculates with motility ≥70% and normal morphology ≥80% were extended into BioXcell extender to which 0 (control), 3, 5, 10 or 15ngmL-1 DHA was added. The supplemented semen samples were incubated at 37°C for 15min for DHA uptake by spermatozoa. Later, samples were cooled for 2h at 5°C and packaged into 0.25-mL straws, frozen in liquid nitrogen for 24h and subsequently thawed for evaluation. Results are presented as percentages ± s.e.m. Supplementation with DHA at 3ngmL-1 significantly improved sperm functional parameters including sperm motility, normal morphology, viability, acrosome integrity and membrane integrity when compared with other supplemented groups and the control. Lipid peroxidation increased as the incorporation of DHA supplementation increased. In conclusion, 3ngmL-1 concentration of DHA resulted in superior quality of frozen-thawed bull spermatozoa and is suggested as the optimum level of DHA to be added into BioXcell extender.

Assessment of blood changes post-challenge with Corynebacterium pseudotuberculosis and its exotoxin (phospholipase D): A comprehensive study in goat

Aim: There is very little information regarding blood changes during the challenge of phospholipase D (PLD) in goats. Therefore, this experiment was conducted to study the changes in blood after the challenge with Corynebacterium pseudotuberculosis and its exotoxin, PLD to fill in the gap of caseous lymphadenitis (CLA) research. Materials and Methods: Twenty-six crossbred Boer goats aged 12-14 months were divided into 3 groups; the first group n=6 was inoculated with 1 ml phosphate buffered solution s.c. as the control. The second group n=10 was inoculated with C. pseudotuberculosis 1 × 109 cfu s.c. The third group n=10 was intravenous injected with PLD 1 ml/20 kg body weight. Serial blood collections were done at 1 h, 3 h, 5 h, 8 h, and 12 h then every 24 h post-inoculation for the first 30 days of the experiment. Subsequently, the blood collection continued twice a week till the end of the experiment (90 days post-challenge). Results: Both C. pseudotuberculosis and PLD treated groups showed significant changes (p<0.05) in red blood cell count, hemoglobin (Hb), packed cell volume, mean corpuscular volume, mean corpuscular Hb concentration, white blood cell count, neutrophils, lymphocytes, monocytes, eosinophils, basophils, globulin, and total plasma proteins. Similarly, both treated groups showed significant changes (p<0.05) in alanine transaminase, alkaline phosphatase, aspartate transaminase, total bilirubin, calcium concentration, creatine phosphokinase, creatinine, gamma-glutamyl transpeptidase, urea concentration, lactate dehydrogenase, prothrombin time, and activated partial thromboplastin time. Conclusion: It concluded that C. pseudotuberculosis and PLD have a negative impact on the goat’s health in general reflected by all those changes recorded in the hemogram, leukogram, and the blood chemistry.

Seroprevalence of Bovine Viral Diarrhea Virus Infection and Associated Risk Factors in Cattle in Selangor, Malaysia

Nurhusien Yimer/ Mohamed Ariff Omar,Jesse Faez Firdaus bin Abdullah,Rosnina Hj. Yusoff,Siti Suri Arshad,Abd Wahid Haron/ / , Kazhal Sarsaif,

Edible bird's nest impact on rats' uterine histomorphology, expressions of genes of growth factors and proliferating cell nuclear antigen, and oxidative stress level

Aim This study aimed to evaluate the effect of edible birds nest (EBN) supplementation on the uteri of rats based on analyses of the morphological and histomorphometric changes, and expressions of epidermal growth factor (EGF) and its receptor (REGF) genes, vascular endothelial growth factor (VEGF), proliferating cell nuclear antigen (PCNA), and steroid receptors. Materials and Methods Twenty-four: Sprague Dawley rats were equally distributed into the following four groups: G1 (control), G2, G3, and G4 represented the groups treated with EBN at graded concentrations of 0, 30, 60, and 120 mg/kg body weight (BW) per day for 8 weeks, respectively. During the experimental period, the BW of each rat was recorded weekly. At the proestrus stage of estrous cycle, blood samples were collected from the hearts of anesthetized rats that were later sacrificed. The uteri were removed for histological and immunohistochemical analyses. Results The EBN-treated groups showed an increase in the weights and lengths of uteri as compared to the control. Results showed that relative to G1 and G2, G3 and G4 exhibited proliferation in their uterine luminal and glandular epithelia and uterine glands, and up-regulated expressions of EGF, REGF, VEGF, PCNA, and progesterone receptor, and estrogen receptor in their uteri. The EBN increased the antioxidant (AO) and total AO capacities and reduced the oxidative stress (OS) levels in non-pregnant rats. Conclusion Findings of this study revealed that EBN promotes proliferation of the uterine structures as evidenced by the upregulation of the expressions of steroid receptors, EGF, REGF, VEGF, and PCNA in the uterus and increased in the plasma concentrations of AO and reduced levels of OS.

Verification of X- and Y-spermatozoa separation by nested polymerase chain reaction (PCR), motility and membrane integrity in bovine

The aim of the present study was to verify the presence of X- and Y-chromosome spermatozoa after separation with swimming speed using oestrus cows vagina mucus, Percoll discontinuous gradient (45 to 90%) and swim-up using TALP medium. The nested polymerase chain reaction (PCR) was used to determine X- and Y-chromosome bearing spermatozoa after separation. The primers for PCR were designed using amelogenin cDNA sequence with 329 and 266 bp for X- and Y-bearing chromosome spermatozoa, respectively. The motility was analyzed using computer assisted sperm analyser, whereas the membrane integrity was analyzed using hypo-osmotic swelling test (HOST). Results were confirm by the absence of single band, either for X- or Y-chromosome. Inversely, the double band indicating that the spermatozoa cannot be separated was observed. The percentage of X-chromosome bearing spermatozoa in the swimming speed using oestrus cows vagina mucus media, Percoll discontinuous gradient and swim up methods were 58.33, 44.33 and 50%, respectively. Statistically, both percentages were significantly different (P<0.001) as compared to the theoretical ratio (50:50). Spermatozoa motility, membrane integrity and concentration before and after separation were also significantly different (P<0.05). This study shows that although swimming speed using oestrus cow’s vagina mucus media may be used to separate X- and Y-chromosome bearing spermatozoa in bulls, the results however, require further investigation.

The Roles of Antioxidants and Fatty Acids in Sperm Cryopreservation

Despite research developments in the area of sperm storage, it has become inevitable to realize a marked reduction in the quality of fresh semen following cryopreservation. As a result, research has continued and will also continue in the future looking forward for a much better and improved methods of sperm cryopreservation along with better understanding of the physical and biochemical challenges that the sperm has to face to survive during freezing. Among the various attempts made to improve the cryopreservation process and subsequently result in superior quality of sperm after thawing include manipulating the composition of semen extenders by addition of exogenous products including antioxidants and fatty acids. While fatty acids are added to strengthening plasma membrane stability, Antioxidants are incorporated to compensate the reduction in the endogenous antioxidants level of seminal plasma due to dilution as well as to combat with the excess reactive oxygen species (ROS) production during freezing. In this chapter, the roles of antioxidants and fatty acids in mammalian sperm cryopreservation, both from endogenous and exogenous perspectives, will be discussed with reference to the latest research findings.

Comparison of the patterns of antral follicular development between hormonally synchronized and natural estrous cycles of non-seasonal, polyestrous goats in the tropics

The effects of estrus synchronization with prostaglandin F(2α) (PGF(2α)) and Controlled Internal Drug Release Device (CIDR) on ensuing antral follicular development were documented and compared to natural estrous cycles of non-seasonal tropical goats. Two to six follicular waves were observed, with the three-follicular wave pattern being most frequently observed (58%), followed by four follicular waves (31.6%) per estrous cycle. There were no significant differences (p>0.05) between the PGF(2α)- or CIDR-synchronized and natural estrous cycles nor between the synchronized and subsequent non-synchronized cycles in terms of the time of ovulation, the duration of inter-ovulatory intervals, daily numbers of antral follicles ≥3mm in diameter, and the number of follicular waves per cycle in the goats of the present study.