• Asian-Australasian Journal of Animal Sciences
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• v.28 no.11
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• pp.1599-1605
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• 2015

Twenty Holstein calves were used to investigate the effects of mannanoligosaccharides (MOS) supplementation in the whole milk on growth performance, faecal score, faecal pH, selected faecal bacterial populations and health during the preweaning period. Healthy calves selected by clinical examination were allocated to one of the two groups (control [CG] and experimental [EG]) at 5 days old. Each group consisted of 5 male and 5 female calves. Each calf in EG was supplemented with 7 g/d of a MOS product (Celmanax) from 5 days to 56 days of age. MOS supplement was mixed with the whole milk once in the morning and administered to the calves in EG via nipple bottle, whereas the calves in CG were fed the whole milk without MOS. Calves were weaned at 56 days of age. The final body weight, average daily weight gain (ADG) and average daily feed intake (ADFI) were statistically similar (p>0.05) but were higher by 3.70%, 6.66%, and 10.97%, respectively, in MOS than in control calves. Feed efficiency (ADG/ADFI) was also similar in two calves group. While faecal scores did not differ on day 5, 7, 14, 21, 28, 42, 49, and 56 between groups, EG had a higher faecal score (p = 0.05) than CG on day 35. Faecal concentration of Lactobacillus was lower (p<0.05) in EG compared with CG. No differences (p>0.05) in faecal concentrations of Bifidobacterium, Clostridium perfringens, and Escherichia coli were found between groups. Although there were no significant differences (p>0.05) in the incidence of diarrhoea, treatment days for diarrhoea and the costs associated with diarrhoea treatments between groups, collectively, the observed reductions in treatment days and the cost of diarrhoea treatments accompanying increases in final body weight, ADG and ADFI for EG may indicate potential benefit of MOS in treatment of diarrhoea.

• Korean Journal of Poultry Science
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• v.40 no.3
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• pp.197-205
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• 2013

This study established a method for preserving chicken primordial germ cells (PGCs) that enables long-term storage in liquid nitrogen ($LN_2$) for preservation of the species. The purpose of this study was to compare the effects of Ethylene Glycol (EG) and Propylene Glycol (PG) on viability of cryopreserved PGCs with vitrification in Korean Native Chicken (Ogye), and to fine should be find or to the optimal protocol for PGCs freezing. One of the important components of cryopreservation process is cryopreservation medium that plays a vital role in preventing cellular injury during freeze-thawing. Cryoprotective agents have been known to improve cell viability after freeze-thawing. PGCs obtained from the germinal gonade of 5.5~6 day (stage 28) chick embryos, using the MACS method were suspended in a freezing medium containing a freezing and protecting agents. Gonads were harvested from stage 28 chick embryos and pooled in groups of 10E embryos, contributing gonads to the cell suspension. The gonadal cells, including PGCs, were then frozen in 1 of the following cryoprotectant treatments: 2.5% EG, 5% EG, 10% EG, 2.5% PG, 5% PG, 10% PG, and 0% cryoprotectant as a control. Effects of exposure to vitrification solution and vitrification, with different concentrations of the cryoprotectant solution, were examined. After freezing and thawing, survival rates of the frozen-thawed PGCs from the 0, 2.5, 5, 10 and 15% EG plus FBS treatment were 44.24%, 64.51%, 85.63%, 80.51% and 73.52% (p<0.05), respectively. The viability of PGCs after freeze-thawing was significantly higher for 10% EG plus FBS treatment than for 10% PG + FBS treatment (p<0.05)(85.63% vs 66.81%). Therefore, these systems may contribute in the improvement of cryopreservation for a scarce species in birds preservation. This study established a method for preserving chicken PGC that enables systematic storage and labeling of cryopreserved PGCs in liquid N at a germplasm repository and ease of entry into a database. In the future, the importance for this new technology is that poultry lines can be conserved while work is being conducted on improving the production of germline chimeras.

• KSBB Journal
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• v.27 no.1
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• pp.51-56
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• 2012

This research was to investigate the polyester preparation using waste ethylene glycol (EG) generated from the wastepaper pretreatment process. Waste EG was obtained from using EG five times repeatedly in the pretreatment of wastepaper. The hydroxyl value of the waste EG was 441 mg KOH/g and its composition was 0.68% cellulose, 6.5% hemicellulose, 6.1% lignin, and 86.7% EG. Maleic acid was used as carboxylic acid. The effect of reaction temperature and time except carboxyl group/hydroxyl group ratio on the crosslinkage of the prepared polyester was marginal. Citric acid, lithium hydroxide and dicumyl peroxide were used as additive or catalyst to enhance the crosslinkage of polyester. Among them, 10% of citric acid was found to be most effective. The crosslinkage was 86% when the polyester was prepared at an optimum condition such as $130^{\circ}C$ and 15 minutes, 1.5 of C/H ratio, and 10% of citric acid, and its insoluble percentage in boiling water for 6 hours was 47%. The weight loss of the prepared polyester was approximately 40% when it was buried in damp soil for 5 months, indicating that it is readily biodegradable. This results can provide some information for future development of wastepaper pretreatment by organic solvent.

• Journal of Animal Science and Technology
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• v.55 no.5
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• pp.417-425
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• 2013

We sought to provide a method for freezing and preserving primordial germ cells, or an avian germ cell of a bird, as a material for developmental engineering or species preservation. The aim of this study was to compare the efficacy of slow freezing with a vitrification method for the cryopreservation of chicken primordial germ cells (PGCs). PGCs obtained from the germinal gonad of day 5.5-6 day (stage 28) cultured chick embryos, using the MACS method, were classified into two groups: slow freezing and vitrification. We examined the viability of PGCs after Cryopreservation. Four freezing methods were compared with each other, including the following: Method 1: The PGCs were frozen by a programmed freezer in a plastic straw, including 2.0 M ethylene glycol (EG) as cryoprotective additive (slow freezing) Method 2: The PGCs were vitrified in a plastic straw, including 8.0 M EG, plus 7% polyvinylpyrrolidone (PVP) (rapid freezing). Method 3: The slow freezing was induced with a cryotube including 2.0 M EG Method 4: The PGCs were frozen in a cryotube including 10% dimethyl suloxide (DMSO) (rapid freezing). After freezing and thawing, survival rates of the frozen-thawed PGCs from Method 1 to 4were 76.4%, 70.6%, 80.5% and 78.1% (p<0.05), respectively. The slow freezing ($-80^{\circ}C$ programmed freezer) method may provide better survival rates of frozen-thawed PGCs than the vitrification method for the cryopreservation of PGCs. Therefore, these systems may contribute to the cryopreservation of a rare avian species.

• Reproductive and Developmental Biology
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• v.39 no.3
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• pp.77-81
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• 2015

This study was carried out to evaluate the effects of embryonic stage and toxicities of cryoprotectant on the rates of survival and development of the cryopreserved mouse early embryo and finally to establish the cryopreservation method of surplus embryos obtained during assisted reproductive technology. Toxicities of two cryoprotectants, dimethyl sulfoxide (DMSO) and ethylene glycol (EG) were investigated using a murine embryo model. Female F-1 mice were stimulated with gonadotropin, induced ovulation with hCG and mated. Two cell embryos were collected and cultured after exposure to either DMSO or EG. Embryo development was evaluated up to the blastocyst stage. The total cell count of blastocysts that were treated with DMSO ($68.1{\pm}24.1$) at the 2-cell stage was significantly lower than that were treated with EG ($81.2{\pm}27.0$) or the control ($99.0{\pm}18.3$) (p<0.001). On comparison of two cryoprotectant treated groups, the DMSO treated group showed a decreased cell count compared with the EG treated group (p<0.05). Both DMSO ($15.4{\pm}1.5$) and EG ($10.2{\pm}1.4$) treated groups showed higher apoptosis rates of cells in the blastocyst compared with the control ($6.1{\pm}0.9$, p<0.0001). In addition, the DMSO treated group showed more apoptotic cells than the EG treated group (p<0.001). The potential toxicity of cryoprotectants was uncovered by prolonged exposure of murine embryos to either DMSO or EG at room temperature. When comparing two cryoprotective agents, EG appeared to be less toxic than DMSO at least in a murine embryo model.

• Journal of Veterinary Clinics
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• v.29 no.1
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• pp.27-32
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• 2012

The purpose of this study was to evaluate survival rates of vitrified mouse blastocysts in various vitrification solutions (cryoprotectants) and apparatuses. The mouse blastocysts were harvested from culture of mouse 2 cell embryo and were divided into three group (i) untreated (control); (ii) exposed to cryoprotectant agents; or (iii) cryopreserved by various vitrification apparatuses. Vitrification solutions are 40% ethylene glycol (EG) + 5.8 mg/mL ficoll + 0.5M sucrose (EFS solution), 3M glycerol + 3M EG (ES solution), 20% EG + 20% dimethyl sulfoxide (ED solution), 3M EG + 1.0 m sucrose (ES solution). Vitrification apparatuses consisted of 5 groups ; closed plastic straw (CPS), electron microscope (EM) grid, cryoloop, open pulled straw (OPS), and glass micropippete in plastic straw (GPS). The survival rates of control were 88.0%. The survival rates of exposed blastocysts in EFS, GE, ED, and ES solutions were 70.8%, 43.5% (P<0.01), 83.3% and 65.2%, respectively. The survival rates of vitrified blastocysts in CPS, EM grid, cryoloop, OPS and GPS were 56.5% (P< 0.01), 72.7%, 83.3%, 60.9% (P<0.05) and 54.2% (P<0.01), respectively. Among the vitrification solutions, the highest survival rate was seen in blastocysts vitrified in EG + DMSO (83.3%). The survival rate was not significantly different from that of the control (88%). Blastocysts cryopreserved with glycerol in all groups had an overall low survival rate of 43.5%. Survival rate of mouse blastocysts between vitrification apparatuses showed higher in cryoloop.

• Journal of Embryo Transfer
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• v.13 no.2
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• pp.173-178
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• 1998

The objective of this study was to evaluate the embryonic development ability and the appearance of blastocysts of bovine in vitro fertilized oocytes cultured in different culture media, and also to evaluate survival rate after thawing of frozen embryos by using 1.5 or 1.8M ethylene glycol(EG) with sucrose or trehalose. Fertilized oocytes were divided into three groups; i ) monolayer of cumulus /granulosa cell prepared by TGM 199+5% calf serum(TGM199), ii)GRlaa+5% CS, iii)SOF+5% CS, and they were cultured after insemination for 9 days, at 39˚C, under 5% $CO_2$ in air, but SOF+5% CS was cultured at 39˚C, under 5% 02, 5% GO2, 99% N2. Blastocysts derived from GRlaa + 5% CS on day 7~8 after insemination were frozen by using 1.5M EG or 1.8M EG with/without 0.2M sucrose or O.1M trehalose. The development rate of blastocysts on day 7 after insemination in SOF+5% CS was significant higher than in TCM199 or CR1aa(P<0.05). The appearance rate of blastocysts on day 7-8 after insemination was higher than in TCM199, when fertilized oocytes were cultured in GRlas or SOF. The survival rate of frozen blastocysts after thawing tended to increase, when blastocysts were frozen by using 1.8M EG with 0.2M sucrose or O.1M trehalose. These results indicated that SOF or CRlaa media with amino acids was superior to TCM199 with monolayer in terms of blastocyst development in culturing of in vitro fertilized bovine nocytes, and sucrose or trehalose was supposed to prevent embryos from the freezing shock.

• Korean Journal of Animal Reproduction
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• v.25 no.4
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• pp.371-379
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• 2001

The objective of this study was to establish an effective cryopreservation method of in vitro-produced bovine embryos. For the vitrification, in virtro-produced embryos at 8-cell, morula and blastocyst stages were exposed to freezing solution containing 5.5 M EG (EG 5.5) for 20 sec, loaded on each containers such as EM grid, OPS and Cryo-loop, and then immediately plunged into liquid nitrogen at -196$^{\circ}C$. Thawed embryos were serially diluted in 0.5, 0.25 and 0.125 M sucrose in m-HPBS, each for 1 min, and cultured in CRlaa medium supplemented with 10% FBS. Significant differences in the rates of re-expanded and hatched embryos were not observed among these embryo containers. The total cell number of expanded blastocyst cultured in vitro after vitrification was examined by Hoechst staining. There were no differences between non-vitrified (180.0 $\pm$ 5.4) and vitrified groups (178.0 $\pm$ 7.5). In addition, when the cellular injuries after vitrification were compared by double staining. There were no significant difference in the ratio of live and dead cells between non-vitrified group (176 : 4) and vitrified group (172 : 6). Therefore, these results suggest that bovine embryos can be cryopreserved easily, effectively and successfully by vitrification using various containers, such as EM grid, OPS or Cryo-loop in the presence of EG 5.5 freezing solution.

• Journal of Life Science
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• v.25 no.6
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• pp.663-672
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• 2015

The study was performed to investigate the effects of enzyme treated garlic (EG) and its natural resources composites on lipid levels in serum and liver of rats fed a high fat diet. Four different types of EG-composite extracts prepared: EG and EG + grape peel (EGG), EG + Persimmon (EGP) and EG + Catechu (EGC) by mixed 9.5:0.5, 9:1 and 8:2 (w/w) ratios, respectively. DPPH and ABTS radical scavenging activity in vitro, show the highest in EG + Catechu (EGC) composite by mixed 8:2 (w/w). EG and EG-composites extracts (8:2, w/w) were administered orally to SD-male rats at a concentration of 2.5 g/kg/day for 5 weeks. Total lipid and cholesterol contents in serum were significantly lower in EGC group than control group, and triglyceride content was the lowest in EGP group by 54.29 mg/dL. HDL-cholesterol contents were significantly higher in EGP and EGC groups. LDL-cholesterol content was lower in EG group than EG-composite groups, and VLDL cholesterol content was the lowest in EGP group. GOT, GPT and ALP activity was significantly lower in EGP group. Total lipid, cholesterol and triglyceride contents in liver were significantly lower in EGP and EGC group than control group. Antioxidant activity in serum was the highest in EGC groups by 50.86%, in liver was the highest in EGP groups. TBARS content in serum and liver was the lowest in EGP group. In these results, we suggest that EGP composites could have hypolipidemic and anti-obesity effects in rats fed a high fat diet.

• Journal of Embryo Transfer
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• v.18 no.3
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• pp.179-186
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• 2003

Human embryonic stem (hES) cell lines have been derived from human blastocysts and are expected to have far-reaching applications in regenerative medicine. The objective of this study is to improve freezing method with less cryo-injuries and best survival rates in hES cells by comparing various vitrification conditions. For the vitrifications, ES cells are exposed to the 4 different cryoprotectants, ethylene glycol (EG), 1,2-propanediol (PROH), EG with dime-thylsulfoxide (DMSO) and EG with PROH. We compared to types of vehicles, such as open pulled straw (OPS) or electron microscopic cooper grids (EM grids). Thawed hES cells were dipped into sequentially holding media with 0.2 M sucrose for 1 min, 0.1 M sucrose for 5 min and holding media for 5 min twice and plated onto a fresh feeder layer. Survival rates of vitrified hES cells were assessed by counting of undifferentiated colonies. It shows high survival rates of hES cells frozen with EG and DMSO (60.8％), or EG and PROH(65.8％) on EM grids better than those of OPS, compared to those frozen with EG alone (2.4％) or PROH alone (0％) alone. The hES cells vitrified with EM grid showed relatively constant colony forming efficiency and survival rates, compared to those of unverified hES cells. The vitrified hES cells retained the normal morphology, alkaline phosphates activity, and the expression of SSEA-3 and 4. Through RT-PCR analysis showed Oct-4 gene expression was down-regulated and embryonic germ layer markers were up-regulated in the vitrified hES cells during spontaneous differentiation. These results show that vitrification method by using EM grid supplemented with EG and PROH in hES cells may be most efficient at present to minimize cyto-toxicity and cellular damage derived by ice crystal formation and furthermore may be employed for clinical application.