Abd Wahid Haron

Effects of basic fibroblast growth factor and leukaemia inhibitory factor on proliferation and short-term culture of human spermatogonial stem cells

In this study, isolated spermatogonial stem cells (SSCs) and Sertoli cells using enzymatic digestion from patients with maturation arrest of spermatogenesis were grown in Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% foetal calf serum in three different groups: (1) SSCs cultured without Sertoli cells (2) SSCs co‐cultured with Sertoli cells (as control group), (3) SSCs co‐cultured with Sertoli cells and adding different concentrations of basic fibroblast growth factor (0.1, 1, 10 ng ml−1) and human leukaemia inhibitory factor (1000, 1200, 1500 unit ml−1) as experimental groups. The assessment of colonies every 10 days during 5‐week cultures showed that in the first group, the average number and diameter of the colonies were significantly lower than in the other groups (P < 0.05). The largest number of colonies was observed in control condition (32.29 ± 9.15) in day 30. The largest diameter of colonies was formed in combination dosages of 1 ng ml−1 basic fibroblast growth factor (bFGF) + 1500 unit ml−1 leukaemia inhibitory factor (LIF) (302.93 ± 37.68) and 10 ng ml−1 bFGF and 1200 unit ml−1 LIF (262.87 ± 35.54) in day 30 respectively. Isolated SSCs were positive for spermatogonial cell markers such as Oct4, Stra8, Piwil2 and Vasa but negative for Nanog. Transplantation technique indicated that hSSCs have good efficiency for colonisation of mouse seminiferous tubules after proliferation in culture system.

The detrimental effects of lead on human and animal health

Lead, a chemical element in the carbon group with symbol Pb (from Latin: Plumbum, meaning “the liquid silver”) and has an atomic number 82 in the periodic table. It was the first element that was characterized by its kind of toxicity. In animal systems, lead (Pb) has been incriminated in a wide spectrum of toxic effects and it is considered one of the persistent ubiquitous heavy metals. Being exposed to this metal could lead to the change of testicular functions in human beings as well as in the wildlife. The lead poising is a real threat to the public health, especially in the developing countries. Accordingly, great efforts on the part of the occupational and public health have been taken to curb the dangers of this metal. Hematopoietic, renal, reproductive, and central nervous system are among the parts of the human body and systems that are vulnerable toward the dangers following exposure to high level of Pb. In this review, we discussed the massive harmful impact that leads acetate toxicity has on the animals and the worrying fact that this harmful toxicant can be found quite easily in the environment and abundance. Highlighting its (Pb) effects on various organs in the biological systems, its economic, as well as scientific importance, with the view to educate the public/professionals who work in this area. In this study, we focus on the current studies and research related to lead toxicity in animals and also to a certain extent toward human as well.

Evaluation of the effects of cryopreservation on viability, proliferation and colony formation of human spermatogonial stem cells in vitro culture

Proliferation of spermatogonial stem cells (SSCs) in vitro system is very important. It can enhance SSCs numbers for success of transplantation and treatment of infertility in cancer patients. In this study, testicular cells that obtained from azoospermia patients (n = 8) by enzymatic digestion were cryopreserved at the beginning and after 2 weeks of culture. Then, frozen‐thawed SSCs were co‐cultured on fresh Sertoli cells (experimental group 1), and frozen‐thawed Sertoli cells (experimental group 2) for another 3 weeks. In control group, fresh SSCs were co‐cultured on fresh Sertoli cells. Viability rate after enzymatic digestion was 93.4%±5.0. Frozen‐thawed testicular cells after 2 weeks of culture had a significantly (P < 0.05) higher percentage of living cells compared to frozen‐thawed testicular cells at the beginning of culture (59.2 ± 7.05 and 46.3 ± 8.40 respectively). The number of colonies in the experimental group 1 was significantly higher than experimental group 2 (19.6 ± 2.8 and 8.33 ± 1.5, respectively, P < 0.05). The diameter of the colonies in the experimental group 1 was significantly higher than control and experimental group 2 (P < 0.05) after 3 weeks of culture (269.7 ± 52.1, 204.34 ± 24.1 and 112.52 ± 23.5 μm, respectively). Cryopreservation technique will raise the possibility of banking SSCs for men who have a cancer‐related illness and waiting for radiotherapy and/or chemotherapy.

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.

Epigenetic modification does not determine the time of POU5F1 transcription activation in cloned bovine embryos

PurposeTo investigate the effect of epigenetic modification on pattern, time and capacity of transcription activation of POU5F1, the key marker of pluripotency, in cloned bovine embryos.MethodsBovine fibroblasts were stably transfected with POU5F1 promoter-driven enhanced green fluorescent protein (EGFP). This provided a visible marker to investigate the effect of post-activation treatment of cloned bovine embryos with trichostatin A (TSA) on time and capacity of POU5F1 expression and its subsequent effect on in vitro development of cloned bovine embryos.ResultsIrrespective of TSA treatment, POU5F1 expression was not detected until 8–16 cell stage, but was detected in both inner cell mass and trophectoderm at the blastocyst stage. TSA treatment significantly increased POU5F1 expression, and the yield and quality of cloned embryo development compared to control.ConclusionThe POU5F1 expression of cloned embryos is strictly controlled by the stage of embryo development and may not be altered by TSA-mediated changes occur in DNA-methylation and histone-acetylation of the genome.

Seroprevalence of orf infection based on IgM antibody detection in sheep and goats from selected small ruminant farms in Malaysia

Orf is an infectious disease that affects the skin of sheep and goats resulting in lesions that reduce animal productivity. This study was aimed to determine the status of orf infection among small ruminants from selected farms in the state of the Selangor based on IgM antibody detection. Serum samples were collected from 90 goats and 90 sheep and subjected to qualitative enzyme-linked immunosorbent assay (ELISA) to measure IgM antibodies followed by chi-square analysis of the data. The result from this study showed that 33 goats (36.7%) and 7 sheep (7.8%) were positive for orf IgM antibodies, indicating higher seroprevalence among goats as compared to sheep. The risk factors such as species, breed, farm location, and history of orf, age, gender, presence of clinical signs, and farm location were shown to significantly affect the seropositivity of IgM antibodies in these species. In conclusion, this study showed that a significant number of goat populations in Selangor, Malaysia, harbor active orf infection in comparison to sheep.

Clinico-pathological Features in Mice Following Oral Exposure to Pasteurella Multocida B: 2

Haemorrhagic septicaemia (HS) is a major cause of losses to livestock production in many countries around the world. In Malaysia, more specifically, the disease yet remains a major constraint to the national industry. However, the pathogenesis of haemorrhagic septicaemia is another scenario in which the limitations still exists. Thus, the present paper provides more information on the pathogenicity and host response dynamics in a mouse model. Our study of experimental nature manipulates P. multocida serotype B:2, the bacterium responsible for the disease in Asia. In this study, sixteen mice (n=16) were divided into two groups (A & B) of 8 mice each group. Animals in group A were inoculated orally with 1.0 ml 10 9 cfu/ml of P. multocida type B while mice in group 2 were challenged orally with 1.0 ml of phosphate buffer saline (PBS). The animals were observed for clinical signs for 5 days. The mice showing severe signs and surviving mice after 5 days of post- inoculation were euthanized using cervical dislocation approach and the organs such as heart, lung, kidney, stomach, spleen, colon and small intestine were collected for microscopic examinations. The result indicated that mice inoculated with the Pasteurella multocida showed significant (p<0.05) severe clinical signs compared to control group.These clinical signs ranged from mild to severe in which most of individuals infected with Pasteurella multocida showed moderate to severe clinical signs of ruffled hair, laboured breathing, eye discharge and responsiveness with mean levels of 2.13±0.64, 1.88±0.99, 1.50±1.20 and 1.88±0.99 respectively in comparison to the control group. Moreover, mortality rate was recorded between 24 to 50 h post-inoculation in the group that challenged with Pasteurella multocida type B: 2. Microscopically, the extent of visceral tissue damages due to the infection was scored. The interested parameters included pulmonary oedema, presence of inflammatory cells, haemorrhage and necrosis. Of these parameters, animals in infected group showed significant (p<0.05) differences in all most all visceral organs. Lungs, liver and kidney were, in particular, the most predominantly affected tissues. Therefore, oral inoculation of P. multocida type B in mice showed significant clinical response and cellular changes.

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.

The re-emerging of orf virus infection: A call for surveillance, vaccination and effective control measures

Orf disease is known to be enzootic among small ruminants in Asia, Africa, and some other parts of the world. The disease caused by orf virus is highly contagious among small ruminant species. Unfortunately, it has been neglected for decades because of the general belief that it only causes a self-limiting disease. On the other hand, in the past it has been reported to cause huge cumulative financial losses in livestock farming. Orf disease is characterized by localized proliferative and persistent skin nodule lesions that can be classified into three forms: generalized, labial and mammary or genitals. It can manifest as benign or malignant types. The later type of orf can remain persistent, often fatal and usually causes a serious outbreak among small ruminant population. Morbidity and mortality rates of orf are higher especially in newly infected kids and lambs. Application of antibiotics together with antipyretic and/or analgesic is highly recommended as a supportive disease management strategy for prevention of subsequent secondary microbial invasion. The presence of various exotic orf virus strains of different origin has been reported in many countries mostly due to poorly controlled cross-border virus transmission. There have been several efforts to develop orf virus vaccines and it was with variable success. The use of conventional vaccines to control orf is a debatable topic due to the concern of short term immunity development. Following re-infection in previously vaccinated animals, it is uncommon to observe the farms involved to experience rapid virus spread and disease outbreak. Meanwhile, cases of zoonosis from infected animals to animal handler are not uncommon. Despite failures to contain the spread of orf virus by the use of conventional vaccines, vaccination of animals with live orf virus is still considered as one of the best choice. The review herein described pertinent issues with regard to the development and use of potential effective vaccines as a control measure against orf virus infection.