This study was conducted to develop a serum-free, defined medium of IVM of pig oocytes. The TCM-199 with supplemented with polyvinylalcohol(PVA), polyvinylpyrrollidone(PVP) and porcine follicular fluid(pFF) were used as basal medium. The effects of the these additives on the rates of maturity and development under in-vitro fertilization and in vitro culture were examined and subsequently considered on the possibilities be sustituted for the bovine serum albumin(BSA). Maturation rate of pig oocytes in IVM media containing PVA(82.4%), pFF(89.4%) and BSA(90.0%) were significantly higher(P<0.05) than that of PVP(78.6%). Cleavage rate after IVF of PVP(64%) was significantly lower(P<0.05) than these of PVA(73%), pFF(77%) and BSA(73%) supplements. in vitro development rates to morulae and blastocyst on PVP(54%) were also significantly lower(P<0.05) than these of the supplements of PVA(63%), pFF(69%) and BSA(65%). In comparison of maturation and fertilization rates of pig oocytes in each supplements, the maturity rates of PVA(82.4%), pFF(89.4%) and BSA(90.0%) were significantly lower(P<0.05) than that of PVP(72.4%) and while, the fertilization rates of pFF(87.1%) and BSA(89.1%) were significantly higher(P<0.05) than these of PVA(78.0%) and PVP(70.6%). It may be concluded that PVA and pFF can be substituted far BSA in medium for culturing pig oocytes; however, it may be considered that PVP were limited to for BSA in the in vitro culture of the embryos.
This research was conducted to investigate the influence of application rate of liming fertilizers on changes in soil chemical properties and growth of 'Red Madness' petunia in plug production. To achieve this, dolomite (DO) with 0, 1.0, 3.5, 8.0 or 13.0 $g{\cdot}L^{-1}$ and calcium carbonate (CC) with 0, 2.0, 2.5, 3.0, 3.5, or 4.0 $g{\cdot}L^{-1}$ were incorporated into peatmoss + vermiculite (1:1, v/v) during the root substrates formulation. The treatments of 3.5 $g{\cdot}L^{-1}$ of DO and 2.5 or 3.0 $gL^{-1}$ of CC had acceptable ranges of pH and EC in soil solution such as 5.6~6.2 and 0.7~1.0 $dS{\cdot}m^{-1}$, respectively. The faster rising of pH was observed in root media containing CC rather than those of DO. This indicates that the solubility of CC is higher than DO. The soil Ca concentrations in all treatments of CC were 1.8 times as high as those of DO. The treatments of 3.5 or 8.0 $g{\cdot}L^{-1}$ of DO had the highest soil Mg concentrations, but all treatments of CC had lower soil Mg concentrations than control treatment indicating that additional application of Mg fertilizers are required. The elevated application rate of DO or CC resulted in the increase of fresh and dry weight. But plant heights were not influenced by application of liming fertilizers. The results of tissue analysis showed that application of DO or CC influenced the $PO_4{^-}P$, Ca and Mg contents, but not influenced the contents of other nutrients such as N, P, Fe, Mn, Zn and Cu.
Uddin, Mohammad Belal;Mukul, Sharif Ahmed;Hossain, Mohammed Kamal
Journal of Forest and Environmental Science
/
v.28
no.4
/
pp.212-219
/
2012
An experiment was set to understand the seedlings growth and nodulation capabilities of five potentially important leguminous agroforestry tree species of Bangladesh in response to application of organic fertilizer. Study was carried out in nursery bed on the seedlings of Acacia mangium Willd., Acacia hybrid, Acacia auriculiformis A. Cunn. ex Benth, Albizia lebbeck (L.) Benth. and Leucaena leucocephala (Lam.) de Wit. after sowing their seeds in polybags filled with different medium. Farmyard manure or decomposed cow dung was applied as the source of organic fertilizer with other potting media. The growth and nodulation performance of seedlings of the selected species were measured three moths after the first seed was emerged. The different soil: organic manure ratios' used during the experiment were, 1:1, 2:1 and 3:1. The effects of organic fertilizer on growth and nodulation were compared to that of seedlings grown in control. The results revealed a positive correlation between seedling growth and the different doses of organic fertilization (i.e. seedling growth was enhanced significantly with different treatments containing higher proportion of organic fertilizer). In few cases a negative effect was observed after applying higher levels of organic manure. It was also apparent that the nodulation in terms of nodule number and size was inhibited significantly by the application of organic fertilizer.
Normal maturation of the mammalian oocytes is prerequisite for the fertilization and the early embryonic development. We have been tested the effects of purine and its de novo synthetic inhibitor, azaserine(Aza) on the maturation of germinal vesicle(GV) and germinal vesicle breakdown(GVBD) mouse oocytes. Denude-immature oocytes were cultivated in the media containing adenosine, guanosine, and/or azaserine, and checked the matruation stage by monitoring the prominent morphological changes. In GV stage oocytes, GV was arrested temporarily by the adenosine(1.0%) and protractedly by the guanosine(65.9%, P<0.001). The regression was increased significantly at the adenosine(90%, P<0.001) but decreased at the guanosine(1.6%, P<0.05). Inhibiting the de novo synthesis of purine, nuclear maturation rate was increase(90.4% : 96.7%), but GV arrest was significantly increased by cotreatment with guanosine(P<0.001). Polar body extraction significantly was increased at the Aza(P<0.05), but not in others. In GVBD oocytes, adenosine itself did not affect GVBD arrest. Guanosine, on the other hand, elevated GVBD arrest rate(P<0.001), but co-treated with Aza, decreased GVBD arrest(P<0.001). Aza increased GVBD arrest rate(20.2%, P<0.05) compared with control. From those results, we know that guanosine shows more prominent effect on the inhibition of nuclear maturation at the GV stage, and of the 1st polar body extrusion at the GVBD stage. Adenosine showed the cytoplasmic toxicity at GV stage oocyte. Our data speculate that cytoplasmic cAMP level is auto-regulated by endogenous adenylate cyclase while GVBD is inhibited by guanosine, since purine toxicity is not observed in the GVBD stage. And it is showed that purine metabolism is concerned with nuclear maturation, that the amounts of purine metabolism is not even during the oocyte maturation.
This study was carried out to investigate on the survival rates and in vitro developmental rates of bisected bovine embryos were by manipulator and micropipette. Bisected embryos were co-cultured in 20% FCS(v/v)+TCM-199 media containing hormones, oviductal epithelial cells and cumulus cells 0 to 72 hrs after bisection. Survival rate and in vitro fertilization rate were defined as development rate on in vitro culture or FDA-test. The results are summarized as follows ; 1. The in vitro developmental rate of biseciton embryos co-cultured in 20% FCS+TCM-199 medium containing PMSG, hCG, PMSG+hCG, hCG+$\beta$-estradiol 0 to 20 hrs and 20 to 40 hrs were 36.7, 26.7, 33.3, 40.0, and 30.0, and 30.0, 33.3, 30.0, 26.7, and 26.7%, respectively. The survival rate of bisection embryos co-cultured in TCM-199 medium containing hormones was significantly higher than that of non co-culture(25.0%). 2. The in vitro developmental rates of bisection embryos co-cultured in 20% FCS+TCM-199 medium containing oviductal epithelial cells 4 to 5 hrs and 20 to 24 hrs were 40.0 and 33.3%, respectively. The survival rate of bisection embryos co-cultured in TCM-199 medium containing oviductal epithelial cells was significantly higher than that of non co-culture(25.0%). 3. The in vitro developmental rates of bisection embryos co-cultured in 20% FCS+TCM-199 medium containing cumulus cells 4 to 5 hrs and 20 to 24 hrs were 43.3 and 36.7%, respectively. The survival rate of bisection embryos co-cultured in TCM-199 medium containing cumulus cells was significantly higher than that of non co-culture (25.0%).
Park Jong-Im;Hwang Woo-Suk;Jo Choong-Ho;Lee Byeong-Chun
Journal of Veterinary Clinics
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v.9
no.1
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pp.323-332
/
1992
The present study was carried out to examine the effect of oviduct epithelium and its conditioned medium on e development of early bovine embryos in vitro. Oocytes obtained from ovarian follicles of slaughtered cows were cultured in TCM199 with 10% fetal calf serum for 22-24hrs and then fertillzed in vitro using frozen-thawed semen treated with BO-caffein, BO-BSA(20mM heparin added). Oviduct epithelium was collected in each stage of the estrus cycle and conditioned medium was the medium in which oviduct epithelium in early luteal stage was cultured. In vitro fertilized bovine embryos of 1~2 cell were co-cultured with oviduct epithelium from different estrus cycles, cultured in conditioned medium, and cultured in rabbit oviduct. The cleavage rates of in vitro fertilized early bovine embryos co-cultured with oviduct epithelial cell from early luteal, luteal and follicular phase of estrus cycle(67.2~70.8%) and cultured in conditioned medium(56.7%) were significantly(p<0.05) higher than that of the control(44.2%) The rate of development to morula or blastocyst stage in oviduct epithelial cell co-culture(15.3~32.5%) from three phase of estrus cycles and conditioned medium(14.5%) were significantly(p<0.05) higher than that of the control(5.2%). The oviduct epithelial cell from early luteal phase gave a significantly( p<0.05) higher rate of development to morula or blastocyst stage than both luteal and follicular phase. The results of in vivo culture in rabbit oviduct of early bovine embryos were 52.1% for the cleavage rate and 26.7% for the rate of development to morula or blastocyst stage.
The influence of hypoxanthine and ovarian steroids on the meiotic maturation process of mouse oocytes was investigated for the qualified application of culture medium in in vitro fertilization(IVF). Mouse oocytes were cultured in hypoxanthine and various ovarian steroids(progesterone, estradiol-17${\beta}$ and testosterone) and their effects on the oocyte maturation had been observed. When mouse oocytes were cultured in the various concentration(1-4mM) of hypoxanthine, meiotic maturation of cumulus cell-enclosed oocytes was inhibited by presence itself, which was a dose-dependent effect in meiotic arrest of mouse oocytes. The presence of progesterone, estradiol-17${\beta}$ and testosterone have made the mouse oocyte mature properly. Meanwhile maturation of cumulus cell-enclosed oocyte was severely inhibited by 3 hoursculture in the media of progesterone supplemented with hypoxanthine. However the continuous presence lasting 24 hours of progesterone even supplemented with hypoxanthine had got rid of the inhibition of oocytes maturation. Not only estradiol-17${\beta}$ supplemented with hypoxanthine but also testosterone supplemented with hypoxanthine exert the severe inhibition of the maturation of cumulus cell-enclosed oocytes for 3-hours culture. However the continuous presence lasting 24 hours of estradiol-17${\beta}$ and testosterone even supplemented with· hypoxanthine had relieved the inhibition of oocytes maturation. These results make us suggest that hypoxanthine inhibits the mouse oocyte maturation, particularly markedly in conjunction with ovarian steroids for short period, which indicated some sort of the synergistic inhibitory retationship between the ovarian steroids and hypoxanthine.
This study was conducted to develop a method for production of nuclear transplant bovine embryos using in vitro-matured (IVM) oocytes and to examine the effect of different conditions of electrofusion on fusion rate and developmental capacity of donor nucleus transplanted to enucleated oocytes. Eight- to sixteen-cell embryos derived from oocytes matured and fertilized in vitro used as donor blastomeres and IVM oocytes were used as recipient oocytes. Oocytes were enucleated immediately after 23-24 h IVM and then reconstituted with a donor blastomere in two different micromanipulation media. Fusion rate and subsequent development of the reconstituted oocytes was compared under the different electric stimuli and recipient oocyte ages. Success rate of enucleation was significantly higher in TCM-199 medium containing FCS than in DPBS. The high fusion rate(75-94%) and development (6.4-14.8%) to morulae and blastocyst (M + B) were obtained from 0.6-0.75 kV/cm DC voltage, although total cleavage was not different among the electric pulses. Most optimal condition of electric stimulation for fusion and development was 1 DC voltage of 0.75 kV/cm, in which 80.5% of oocytes were fused, 80.0% and 31.7% of which was cleaved and developed to M + B, respectively. No M + B was obtained from 1.2 kV/cm DC voltage regardless of pulse frequency. Recipint oocyte age at electrofusion greatly affected the cleavage and subsequent development to M + B, showing high rate at 40-41 h oocyte maturation. These results suggest that a suitable condition of electrofusion for donor nuclei derived from IVF may be 1-2 DC pulses of 0.7 kV/cm for $70{\mu}sec$ and that processing of a transplanted nucleus in IVM oocytes may be affected by maturation age of recipient oocytes.
Cultural hybridization and global cross-fertilization, begun since the 1990s, mean the hybrid mixture of the low-class popular arts and the high-class fine arts and have found expression in intertextuality by means of the thinking system of de-constructuralist post-modernism. This study was intended to investigate the artistic intertextuality between fashion and art frontiere, especially between such fields as 'animation', 'music', and 'performing arts' and 'fashion', which shows the greatest characteristic of the phenomenon that the non-mainstream culture flows into the mainstream culture. It also aimed to investigate the intertextuality between 'technology' and 'fashion' as the delivery room of the youth culture and copycat culture due to the benefit of technologies since the 20th-century digital revolution. Animation as the neo-pop art with the popular code coming to the forefront in the 21th century integrated high-class fashion and low-class fashion into one through the combination of fashion and humor, and the musical element referring to the social difference of the cultural field and the social strata is becoming the 21th-century fashion icon through its fusion with the pictorial tendency along with the leisure-time life of the non-mainstream strata. In terms of intertextuality in performing arts, fashion style performs an important role in the presentation of performing arts and since the de-construction of the fin-de-ciecle form and its fusion with media have taken place, fashion works become and element of the origin that has an influence on a series of film, dance, dramatic elements and the like. The paradigm of technology made it socially and culturally possible to achieve the architecture of clothing system necessary for fashionable technical clothing by allowing the possibility of imitation, the function of technology to form popular culture. The intertextual tendency in the 21th-century fashion began from the de-constructive phenomenon of existing norms and now takes the multicultural character of surpassing a certain domain or concept. And it positions itself as the total art of spearheading the low-class culture and the non-mainstream culture as the hybrid domain of mainstream and non-mainstream cultures or high-class and low-class cultures.
The final biomass yields of 16 marine phytoplankton clones were measured in media with different levels of iron and phosphorus concentrations. The biomass yields of oceanic clones were either only slightly limited or not limited by iron, and those of coastal clones were severely limited by iron in all photogenetic groups other than cyanobacteria. By contrast oceanic cyanobacteria clones as well as coastal clones required higher iron concentrations: minimum concentrations of iron addition for detectable growth of the Synchronous species were estimated to fall between $10^{-9}{\;}and{\;}10^{-8}{\;}M$. Not only the habitat differences (oceanic-coastal trends) but also the photogenetic differences of the oceanic phytoplankton species in the response to the iron enrichments deserve very careful attention before ocean iron fertilization.
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