• Food Science and Preservation
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• v.30 no.3
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• pp.395-404
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• 2023

Essential oil (EO) was extracted from the peel of Citrus hystrix DC. originating from Tinh Bien, An Giang province (Vietnam), using steam distillation. The study aimed to determine some physicochemical properties of Citrus hystrix peel EO (ChpEO), including the acid value (AV), saponification value (SV), ester value (EV), density, specific gravity, and freezing point. The chemical composition was also analyzed by gas chromatography-mass spectrometry (GC-MS). Compounds like β-pinene (30.19%), D-limonene (22.15%), and sabinene (21.37%), with antioxidant and antibacterial properties, had a relatively high content. The EO was also capable of inhibiting the growth of both Gram-positive and Gram-negative bacteria, including Pseudomonas aeruginosa (ATCC 27853), Staphylococcus aureus (ATCC 25923), Salmonella typhimurium (ATCC 13311), and Bacillus cereus (ATCC 11778) specifically.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.11
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• pp.1527-1533
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• 2001

Garole is a prolific, rare, less known and small size Indian sheep breed found in low and humid Sunderban region of West Bengal. Although information on stored Garole ram liquid semen upto 24 h is available, but there is a need to further investigate the short-term and long-term preservability of Garole ram semen for extensive utilization of this valuable germplasm by artificial insemination. The aim of the present study was to apply computer-assisted sperm analysis technique for assessing the motion characteristics of Garole ram semen stored (i) in liquid state at refrigeration temperature for short-term preservation upto 48 h and (ii) in frozen state at $-196^{\circ}C$ for long-term preservation after packaging in mini straws. Short-term preservation had a significant effect on motility (p<0.01) as the motility progressively decreased from 90.1% at 0 h to 85.5% and 73.2% after 24 and 48 h of storage, respectively. Although the decline in rapid moving sperms was also significant (p<0.01) on storage but the decrease was more pronounced at 48 h as compared to 24 h of storage period. Storage of chilled semen had also a significant effect on % linearity (p<0.05), % straightness (p<0.01), sperm velocities (p<0.01), amplitude of lateral head displacement (p<0.01) and beat frequency (pO.Ol) of spermatozoa. The replication had a significant effect for all the variables except average path and straight line velocity. However, the interactions of short-term storage and replication were non-significant for most of the variables except % of medium moving sperms, sperm velocities and beat frequency. On long-term preservation of Garole ram spermatozoa under controlled conditions the mean post-thaw recovery of 70.4 and 71.4% motile spermatozoa was achieved having 48.8 and 48.9% of rapidly motile spermatozoa, respectively in both the replicates. The effect of replication on cryopreservation was significant (p<0.05) on amplitude of lateral head displacement and beat frequency, but there was no significant effect on motility, rapidly motile spermatozoa, linearity, straightness and sperm velocities of frozen-thawed spermatozoa. It can be concluded from these results that an average 70% motility can be achieved on storage of Garole ram semen in chilled liquid state upto 48 h or in liquid nitrogen after freezing under controlled conditions in straws. However, further studies are required to evaluate the fertility of short-term and long-term preserved Garole ram semen for extensive use of this prolific sheep breed.

• Journal of Embryo Transfer
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• v.23 no.3
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• pp.223-227
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• 2008

The cryopreservation of Hanwoo embryos has become an integral part of assisted reproduction in animal. The objective of this study was to assess the effect of The objectives of this study were: (1) to evaluate the influence of bovine embryo developmental stage on in vitro embryo development after freezing, (2) to study the efficiency compared with conventional freezed embryos at different embryo source. For conventional slow-freezing, day 7 or 8 expanded blastocysts were collected. The standard freezing medium was 1.8 M ethylene glycol (EG). Embryos were equilibrated in 1.8 Methylene glycol(EG) with 0.1 M sucrose in Dulbecco's phosphate-buffered saline (D-PBS) supplemented with 0.5% bovine serum albumin. Embryos were then loaded individually into 0.25 ml-straw and placed directly into cooling chamber of programmable freezer precooled to $-7^{\circ}C$, after 2 min, the straw was seeded, maintained at $-7^{\circ}C$ for 8 min, and then cooled to $-35^{\circ}C$ at $0.3^{\circ}C$/min, plunged and stored in liquid nitrogen for at least 3 days. For thawing, the straw containing embryos were warmed in air for 10 see and exposed to $37^{\circ}C$ water for 20 sec. Straws were then removed from $37^{\circ}C$ water. Rates of blastocyst survive and hatched were evaluated at 12 to 48h post-warming. The re-expansion and hatched rates of morula embryos were significantly lower than those obtained for blastocysts and expansion blastocysts (31.6%, 10.5% vs, 68.9%, 22.2% vs, 73.7%, 53.6%, respectively). No differences in re-expansion rates were found between in vivo and in vitro blastocysts. whereas hatched rates was significantly higher (51.2%) in vivo compared with in vitro embryos (18.6%). in conclusion, demonstrate that conventional freezing can be used successfully in cryopreservation of in vitro and in vivo bovine embryos, and that it might be considered for use in commercial programs and embryo preservation.

• Journal of Embryo Transfer
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• v.27 no.1
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• pp.29-35
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• 2012

The purpose of this study was to examine the effect of growing stages of the Korean Native Striped Bull (KNSB) on the freezability and fertility of frozen-thawed semen. First, we investigated the total motility (TM) and progressive motility (PM) according to the diluent used for semen freezing. Second, we examined the effect of the age of KNSB on semen volume, TM and PM of fresh and frozen-thawed semen. Third, we examined the effect of frozen semen from the different age of KNSB on the $in-vitro$ fertilization rate, and the artificial insemination pregnancy rate. The diluents used in this experiment were Triladyl$^{(R)}$ and Tris-egg yolk extender (EYE). Semen was collected from 5 KNSB in the growing stage (15 months) and 5 adult KNSB (36 months). When Triladyl or Tris-EYE extender was used for semen freezing, there was no difference of the mean TM and the mean PM. However, the mean TM was significantly higher in Bull No. 1885 than Bull No. 4283 ($p$ <0.05). The mean volume of semen collected from the 15-month-old bulls (2.3 ml) was significantly lower ($p$ <0.05) than that from the 36-month-old bulls (5.0 ml). The mean semen concentration was similar for the 15-month-old ($2.1{\times}10^9$ spermatozoa/ml) and 36-month-old ($1.8{\times}10^9$ spermatozoa/ml) bulls. For the 15-month-old and 36-month-old bulls, the mean TM of fresh semen were 93.7% and 88.3%, respectively, and the mean PM were 97.0% and 88.3%, respectively; the 15-month-old bulls showed a particularly high PM ($p$ <0.05). For the 15-month-old and 36-month-old bulls, the mean TM (56.0% and 58.0%, respectively) and the mean PM (64.0% and 70.7%, respectively) of frozen-thawed semen did not differ. The development rates of embryos after $in-vitro$ fertilization and the pregnancy rate after artificial insemination using frozen-thawed semen did not differ according to the bull's age. In summary, semen volume differed according to the bull's age, but semen concentration and survival rate, the $in-vitro$ fertilization rate, and the pregnancy rate did not differ according to the stripe bull's age. Accordingly, semen from bulls in the growing stage can be collected and frozen for the preservation and multiplication of rare livestock.

• Clinical and Experimental Reproductive Medicine
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• v.43 no.2
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• pp.119-125
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• 2016

Objective: The aim of this study was to report a case series of in vitro matured (IVM) oocyte freezing in gynecologic cancer patients undergoing radical surgery under time constraints as an option for fertility preservation (FP). Methods: Case series report. University-based in vitro fertilization center. Six gynecologic cancer patients who were scheduled to undergo radical surgery the next day were referred for FP. The patients had endometrial (n=2), ovarian (n=3), and double primary endometrial and ovarian (n=1) cancer. Ex vivo retrieval of immature oocytes from macroscopically normal ovarian tissue was followed by mature oocyte freezing after IVM or embryo freezing with intracytoplasmic sperm injection. Results: A total of 53 oocytes were retrieved from five patients, with a mean of 10.6 oocytes per patient. After IVM, a total of 36 mature oocytes were obtained, demonstrating a 67.9% maturation rate. With regard to the ovarian cancer patients, seven IVM oocytes were frozen from patient 3, who had stage IC cancer, whereas one IVM oocyte was frozen from patient 4, who had stage IV cancer despite being of a similar age. With regard to the endometrial cancer patients, 15 IVM oocytes from patient 1 were frozen. Five embryos were frozen after the fertilization of IVM oocytes from patient 6. Conclusion: Immature oocytes can be successfully retrieved ex vivo from macroscopically normal ovarian tissue before radical surgery. IVM oocyte freezing provides a possible FP option in patients with advanced-stage endometrial or ovarian cancer without the risk of cancer cell spillage or time delays.

• Food Science and Preservation
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• v.20 no.3
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• pp.296-303
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• 2013

This study was conducted to assess the possibility of preserving frozen vegetables (Aster scaber, soybean sprouts, Chinese cabbage, green pumpkin, and Welsh onion) for a long period and of using them after such storage by measuring changes in quality due to their preservation. Various freshly harvested vegetables were blanched under optimal conditions (that were determined in a preliminary experiment), quick-frozen at $-40^{\circ}C$ for 24 h, and stored at $-20^{\circ}C$. The change in the chromaticity of the frozen A. scaber, soybean sprouts, Chinese cabbage, green pumpkin, and Welsh onion did not vary. The hardness of the frozen A. scaber, green pumpkin, Chinese cabbage and Welsh onion did not change during the preservation period, whereas the hardness of the cotyledon and hypocotyl of the soybean sprouts significantly increased on the sixth month of their storage. The total bacterial counts of the A. scaber during the preservation period remained constant at $10^3$ CFU/g, whereas those of soybean sprouts, Chinese cabbage, green pumpkin, and Welsh onion decreased slightly to $10^2$ CFU/g. Coliform was not detected in any of the samples. The sensory evaluation showed that the preservation period that was used in this study did not significantly affect the marketability of the frozen vegetables. Therefore, it was considered that A. scaber, soybean sprouts, Chinese cabbage, green pumpkin, and Welsh onion can be safely preserved by freeze-storing them for up to 12 months.

• Development and Reproduction
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• v.11 no.2
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• pp.79-84
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• 2007

The freezing susceptibilities of two larval stages (D-shaped and umbo) of the pearl oyster (Pinctada fucata martensii) were evaluated by the electron microscopy (light, transmission electron and scanning electron). The morphological shapes were examined from each pre-frozen or frozen-thawed stage of the cryopreserved larvae in liquid nitrogen by using the cryoprotectant, dimethyl sulfoxide ($Me_2SO$) mixed with sucrose. Although a portion of the shell was damaged, the hinge and prodissoconch were intact and clearly visible after preservation in liquid nitrogen. In addition, the cytoplasm of the frozen-thawed larvae maintained the normal organelle integrities, e.g., endoplasmic reticula, lipid droplets, mitochondria, nucleus and microvilli. However, some of the frozen-thawed larvae showed irregularly arranged cilia, rough shell surfaces and round-lumped cilium heads. These results indicate that P. fucata martensii larvae are susceptible to freezing, at least at those two critical developmental stages (D-shaped and umbo), and suggest a new industrial investigation including reduction method of cell injury for preserving microbial starter cultures need to be developed.

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.295-300
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• 2011

The objective of this study was to evaluate efficiency of methyl-beta-cyclodextrin (MBCD) in the sperm preservation of bull. For this study, the freezing of diluted semen were added with Triladyl containing 20% egg-yolk and/or 0, 1, 5, 10 and 20 mM MBCD before freezing process. Analysis of viability in frozen-thawed sperm was estimated by SYBR14/PI double stain, hypoosmotic swelling test(HOST) and acrosome damage with FITC-PNA, and mitochondria activation with Rhodamin123 by flow-cytometry. The sperm viability was significantly in 0 mM and 5 mM concentrations of MBCD than other groups (p<0.05). However, the HOST was significantly lower at 20 mM concentration of MBCD than other concentrations (p<0.05). In addition, acrosome damage and mitochondria activation rates were significantly lower at 20 mM concentration of MBCD than other groups (p<0.05). In conclusion, the viability of sperm was not significantly different among concentrations of MBCD 0, 5 and 10 mM, but MBCD 20 mM was significantly lower than other groups. In addition, as concentrations of MBCD was high, HOST, acrosome damage and mitochondria activation rates had a negative effect in bull sperm.

• The Journal of The Korea Institute of Intelligent Transport Systems
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• v.6 no.3
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• pp.33-44
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• 2007

Prohibition of lane changes in bridges and tunnels have been many problems in throughputs of expressways caused by heavy vehicles and slow-moving traffics. Nevertheless, those are constructed actively by the general trends, which are preservation of environment and ecosystem are more important, because mountainous districts are about 70% across the whole extent of Korea. In this paper, the proper design standards for permission of lane changes in bridges and tunnels classified into structure, safe, and driver's conveniences are suggested as follows. 1. Right shoulder should have more than 2.5m in bridges and tunnels. 2. Sufficient equipments of guidance like as directional signs, fingerposts, variable message signs, and markings should be established to smooth and safe lane changes of drivers. 3. Snow melting systems should be established in bridges worried about freezing. 4. Tunnels must be not only satisfied standards for prevention of disasters (2004.11) and lighting rules (KSA 3703), but also established anti-freezing facilities in entrance and exit. 5. The drivers should have honed on their car lights.

• Reproductive and Developmental Biology
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• v.30 no.4
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• pp.279-285
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• 2006

Vitrification has been suggested to be an effective method for the cryopreservation of human ES cells. However, the efficiency of vitrification with different vehicles remains a matter of ongoing controversy. The objective of this study was to assess the efficiency of cryopreservation in human ES cells by vitrification using different vehicles. A human ES cell line and a variety of vehicles, including micro-droplet (MD), open-pulled straw (OPS) and electron microscopic grid (EM-grid), were employed in an attempt to assess vitrification efficiency. In order to evaluate the survivability and the undifferentiated state of the post-vitrified human ES cells, we conducted alkaline phosphatase staining and characterization via both RT-PCR and immunofluorescence assays. The survival rates of the post-vitrified human ES cells using MD, OPS and EM-grid were determined to be 61.5%, 66.6% and 53.8%, respectively. There also exist significant differences between slow-freezing and vitrification (p<0.01). However, no significant differences were detected between the vehicle types. Finally, the pluripotency of human ES cells after thawing was verified by teratoma formation. Cryopreservation using vitrification is more effective than slow-freezing, and the efficiency of vehicles proved effective with regard to the preservation of human ES cells.