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Articles by Alexandru RUSU
Total Records ( 3 ) for Alexandru RUSU
  Marius ZAHAN , Vasile MICLEA , Florin GHIURU , Iulian ROMAN , Alexandru RUSU , Ileana MICLEA and Manuel MIHAILESCU
  Despite the new researches in boar semen cryopreservation, application of frozenthawed (FT) semen in commercial artificial insemination (AI) programs is still limited. The main sources of semen used in AI are semen preserved on room temperature (RT) and sometimes fresh (F) semen. However, the use of male germplasm from genebank on some rare breeds is connected with the ability to use frozen semen. The aim of this study was to evaluate the effect of semen preservation method and especially the influence of freezing on Red Mangalitsa and Bazna boar semen cryopreservation. Six sexually mature boars from Red Mangalitsa and Bazna rare breeds of the Faculty of Animal Sciences and Biotechnology, USAMV Cluj-Napoca, were used in this study. Semen was collected using the gloved-hand method, evaluated for conventional semen characteristics and extended in Beltsville Thawing Solution (BTS). Preservation was made by RT and by frozen using a modified method described by Westendorf et al. (1975), in lactose, egg yolk, glycerol and Orvus ES Paste extender (LEYGO). The F, RT and FT sperm was evaluated by Eosin-nigrosin viability staining, acrosomal integrity shown by normal apical ridge (NAR) and plasma membrane integrity by hypo-osmotic swelling test (HOST) in order to establish sperm quality. Because it is known that the life of sperm is short in utero after thawing, the FT sperm quality was also determined by thermoresistance test (motility at 10, 30, 60, 120 and 240 minutes). The results were statistically analyzed using ANOVA. The first parameter affected by freezing was viability, but the state of the acrosome has shown significant differences between the preservation methods on both breeds. In comparison with F and RT semen the value of NAR was halved. In addition, membrane integrity spermatozoa (HOST) was significantly lower after the FT procedure compared to F and RT. Thermoresistance test has shown a good motility until 120 minutes, with the best value at 30 minutes, but decreased dramatically at 240 minutes. However, this results correlated with the reducing insemination-to-ovulation interval could improve the fertility after AI with Red Mangalitsa and Bazna breeds’ semen cryopreservation
  Iulian ROMAN , Vasile MICLEA , Andrea HETTIG , Marius ZAHAN , Ileana MICLEA , Florin VARO-GHIURU and Alexandru RUSU
  Supplementation of a chemical defined in vitro maturation (IVM) media with some amino acids was examined in the terms of oocyte maturation and fecundation rate following intracytoplasmic sperm injection (ICSI). Addition of glutamine resulted in a higher (p<0.05) in vitro maturation rate, compared to the total absence of amino acids in media, but in terms of fecundation rate, none of the amino acids used increased the fecundation rate in our research.
  Andrea HETTIG , Vasile MICLEA , Marius ZAHAN , Iulian ROMAN , Florin VARO-GHIURU and Alexandru RUSU
  The aim of this study was to establish a protocol with a higher number of viable oocytes after the vitrification procedure. The oocytes were collected by the follicular punction method from swine prepuberal ovaries. A total of 1792 immature oocytes were cryopreserved stepwise in previtrification (PV) and vitrification (VM) media, containing ethylene glycol (EG) permeating agent, combined with trehalose a non-permeating agent. The cells were preserved in three experimental ways, with three different concentration of cryoprotectant in the VM respectively 30, 40 and 45% EG. The exposure time of the cells to the cryoprotectants was the same in each experimental design, 4 minutes in the PV and 40 seconds in the VM. The concentration of the trehalose and the EG in the PM were remained the same. The vitrification procedure was performed by the superfine open pull straw method. For each experiment, there were 5 repetitions. After tawing the cells were stained with fluoresceine diacetate (FDA) and propidium iodide (PI), and there were exposed to the microscope UV light to test their viability. The viable oocytes had a florescent green color because of the FDA staining, which binds the esterase present only in the viable cells. The PI penetrates only dead cells, with destroyed membrane, binding the DNA, and giving them a red color. The results show an increase of percentage of viable cells once the concentration of the cryoprotectant is increased. With a 30% of EG in the VM, the medium percentage of viable cells is 10,26%, with a standard deviation of 1, 06. For 40% EG in the VM, the median is 33, 81%, with a standard deviation of 0,96, and 46,30% for a concentration of 45% EG in the VM with a standard deviation of 0,50. The statistical analysis shows significant differences (p< o, ooo1) between the experiments. Vitrification is the solidification of a solution, glass formation, at low temperatures without ice crystal formation. The phenomenon can be regarded as an extreme increase of viscosity and requires either rapid cooling rates or the use of cryoprotectant solutions, which depress ice crystal formation and increase viscosity at low temperatures. (Vajta G., 2000). These results sustain the theory according to which the higher the cryoprotectant is, the better the result is but limited by the exposure time.
 
 
 
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