• Food Science and Preservation
• /
• v.5 no.4
• /
• pp.326-330
• /
• 1998

Freeze storage technique is widely used for food processing to keep freshness and quality of the product. This technique was applied to fresh, unshelled groundnut to develop a new type of product which could maintain fresh taste and nutritive values even after several months of storage. The groundnut varieties, Daepungtangkong, Daekwangtangkong and Sindaekwangtangkong were grown at the experimental field of NCES in 1996. Immatured pods or groundnut were harvested around 20 to 3o days before full maturity, washed, and steamed at 100$^{\circ}C$ for 5 min. to stop enzyme activity. After vacuum packing (at -760mmHg for 10 min.) with 0.08mm polyvinyldichloride film, the pods were immediately frozen at -70$^{\circ}C$ for 24h and transfered at -20$^{\circ}C$ for long-term storage. Physico-chemical properties of frozen vegetable groundnut were investigated at 2 months after storage and compared to those of conventionally dried groundnut. After 2 months storage, the thawed kernels were very palatable with softness and fresh taste. Acid value and hardness (measured as the compression force on the probe of a texture analyzer) were much lower in frozen vegetable groundnut than those in the air-dried ones. Presence of free sugars is one of the important factors affecting groundnut taste, and the free sugar contents were considerably decreased in the frozen vegetable groundnut compared to freshly harvested groundnut. But in dried groundnut no free sugar was detected.

• Journal of Embryo Transfer
• /
• v.12 no.1
• /
• pp.11-20
• /
• 1997

The influence of cryopreservation of donor embryos on the in vitro developmental potential in the nuclear transplant rabbit embryos was evaluated. The embryos of 16-cell stage were collected and cryopreserved with EFS solution by vitrification method. The frozen embryos were thawed and synchronized to S and G$_1$ phase of 32-cell stage. The recipient/ cytoplasms were obtained by removing the first polar body and chromosome mass from the oocytes collected by non-disruptive microsurgery procedure. The separated S and G$_1$ phase blastomeres of 32-cell stage were injected into enucleated recipient cytoplasms by micromanipulation. After culture until 20 hrs post-hCG injection, the nuclear transplant oocytes were electrofused and activated by electrical stimulation. The fused nuclear transplant embryos were co-cultured with rabbit oviduct epithelial cells. After in vitro culture for 120 hrs, the nuclear transplant embryos developed to blastocyst stage were stained with Hoechst 33342 dye and their blastomeres were counted. The electrofusion rate was significantly (P<0.05) reduced in the frozen nuclear donor,compared with fresh donor nuclei as 80.0 vs 62.8% in S phase and 81.7 vs 64.8% in G$_1$phase, respectivley. The in vitro developmental rate to blastocyst stage with the S and G$_1$phase of fresh embryos(26.3 and 61.1%, respectively) was found significantly (P<0.05) higher, compared to the S and G]phase of frozen embryos(11.9 and 34.6%, respectively). When frozen as well as fresh donor embryos were synchronized to G$_1$ phase, the in vitro developmental rate to blastocyst stage was significantly (P<0.05) higher, compared with S phase donor nuclei. The cell counts of nuclear transplant embryos developed to blastosyst stage were significantly (P<0.05) more in G$_1$ phase of fresh or frozen embryos (180.1 and 125.7 cells, respectively), compared with S phase nuclear donor (145.1 and 103.7 cells, respectively). From the above results it was concluded that the rabbit embryos cryo- preserved by vitrification might be available as nuclear donor, though the developmentalpotential and cell counts of nuclear transplant rabbit embryos were decreased significantly.

• Journal of Animal Science and Technology
• /
• v.65 no.2
• /
• pp.401-411
• /
• 2023

Many studies have been conducted to improve technology for semen cryopreservation in pigs. However, computer-assisted analysis of sperm motility and morphology is insufficient to predict the molecular function of frozen-thawed semen. More accurate expression patterns of boar sperm proteins may be derived using the isobaric tags for relative and absolute quantification (iTRAQ) technique. In this study, the iTRAQ-labeling system was coupled with liquid chromatography tandem-mass spectrometry (LC-MS/MS) analysis to identify differentially expressed CM10-fractionated proteins between fresh and frozen-thawed boar semen. A total of 76 protein types were identified to be differentially expressed, among which 9 and 67 proteins showed higher and lower expression in frozen-thawed than in fresh sperm samples, respectively. The classified functions of these proteins included oxidative phosphorylation, mitochondrial inner membrane and matrix, and pyruvate metabolic processes, which are involved in adenosine triphosphate (ATP) synthesis; and sperm flagellum and motile cilium, which are involved in sperm tail structure. These results suggest a possible network of biomarkers associated with survival after the cryopreservation of Duroc boar semen.

• Food Science of Animal Resources
• /
• v.38 no.5
• /
• pp.959-969
• /
• 2018

This study investigated the effects of deep freezing and storage temperature ($-50^{\circ}C$, $-60^{\circ}C$, and $-80^{\circ}C$) on the quality and freshness of lamb. To compare the qualities of deep frozen and stored lamb, fresh control and normal freezing conditions ($-18^{\circ}C$) were adopted. As quality and freshness parameters, drip loss (thawing loss and cooking loss), water-holding capacity (WHC), texture profile analysis (TPA), thiobarbituric acid reactive substances (TBARS), and total volatile basic nitrogen (TVBN) were evaluated during 5 months of storage. Temperature influenced the drip loss and WHC, and deep freezing minimized the moisture loss during frozen storage compared to the normal freezing condition. Lamb frozen and stored at deep freezing temperature showed better tenderness than that stored in normal freezing conditions. In particular, lamb frozen at lower than $-60^{\circ}C$ exhibited fresh lamb-like tenderness. Regardless of temperature, evidence of lipid oxidation was not found in any frozen lamb after 5 months, while TVBN was dependent on the applied temperature. Therefore, this study demonstrated that deep freezing could potentially be used to maintain freshness of lamb for 5 months. From the quality and economic aspects, the freezing and storage condition of $-60^{\circ}C$ is estimated as the optimum condition for frozen lamb.

• Journal of Embryo Transfer
• /
• v.5 no.2
• /
• pp.38-44
• /
• 1990

This study was carried out to produce superior dairy cattle by embryo transfer. Seven dairy cows were superovulated with divided injection of FSH 4Omg for 5 days started on day 9 to 14 of the estrus cycle and injection of PGF$_2$$\alpha$ 45mg on day 4 of FSH injection. Donor cows were flushed to collect embryos on day 7 or 8 of the estrus cycle. Fresh embryos collected were transferred to synchronized dairy recipients or frozen using glycerol 3 step method to he equilibrated. And 35 embryos which were frozen using glycerol 6 step method were imported from U.S.A. After glycerol dilution of frozen embryos was done by reverse density during freezing. frozen-thawed embryos were transferred to synchronized dairy or beef recipients. The results obtained were as follows; 1. Total of 24 embryos were collected from 7 donor cows flushed and transferable embryos were 18 (75.0%). 2. Among 24 embryos. morula, early blastocyst, blastocyst, expanded blastocyst and unfertilized ova were 3 (12.5%), 1 (4.2%), 10 (41.6%), 4 (16.7%) and 6 (25.0%), respectively. 3. Heat inducing rate after 1st and 2nd injections of PGF$_2$$\alpha$ in Holstein and beef cattle was 83.3% and 71.4% and 62.5% and 69.2%, respectively. 4. Among 56 recipients, 23 head were pregnant (41.1%). The pregnancy rate of fresh embryos was 50.0% (1/2 heads) and the pregnancy rate of frozen embryos which were frozen using glycerol 3 step and using glycerol 6 step imported from U.S.A. was 52.6%(l0/19 heads) and 34.3%(12/35 heads), respectively. 5. The pregnancy rate of blastocyst (60.0%) was higher than that of morula (39.0%), early blastocyst (25.0%) and expanded blastocyst (0%). 6. The pregnancy rate of grade I embryos (52.2%) was higher than that of grade 2 (34.6%) and grade 3 (28.6%). 7. The pregnancy rate according to synchrony of recipient with donor was higher in simultaneous recipient (55.0%) and +l2hrs' (53.8%) than -24hrs' (23.5%), -l2hrs' (20.0%) and +24hrs' (0%).

• Korean Journal of Animal Reproduction
• /
• v.13 no.1
• /
• pp.18-25
• /
• 1989

This study was carried out to investigate the general semen characteristics of the Korean native goat and the effect of temperature, incubation time, dilution rate, freezing rate and glycerol concentration on motility and NAR (normal apical ridge) acrosome of fresh and frozen Korean native goat spermatozoa. 1. Average semen volume per ejaculate, motility, concentration and pH of fresh Korean native goat spermatozoa were 0.19${\pm}$0.09 ml, 94.5${\pm}$0.47%, 26.17${\times}$108${\pm}$1.68/ml and 6.63${\pm}$0.18, respectively. 2. Motility and NAR acrosome of fresh spermatozoa during incubation were higher at 22$^{\circ}C$ than at 5$^{\circ}C$ or 37$^{\circ}C$(P<.01). 3. Motility and NAR acrosome of spermatozoa diluted 1:4 during incubation were higher at 22$^{\circ}C$ than at 5$^{\circ}C$ or 37$^{\circ}C$(P<.01). 4. Motility and NAR acrosome of spermatozoa during incubation were higher for samples diluted 1:1, 1:2, or 1:4 than for samples diluted 1:6(P<.01). 5. Motility and NAR acrosome of post-thaw spermatozoa were higher at freezing rate of 12$^{\circ}C$/min than at freezing rate of 1$^{\circ}C$/min or 24$^{\circ}C$/min when glycerol concentration was 9%(P<.01).

• Food Science of Animal Resources
• /
• v.40 no.4
• /
• pp.601-612
• /
• 2020

The study assessed the stability for fresh beef patties with the inclusion of clove extract (CE) as a natural antioxidant in comparison to butylated hydroxytoluene (BHT) and ascorbic acid (AA) at frozen storage. Four different patties were made dependent on the added antioxidants: control (added no antioxidants), added with 0.02% BHT, 0.05% AA, and 0.1% CE. Inclusion of BHT, AA, and CE resulted in a significant reduction of thiobarbituric acid reactive substances (TBARS) and hue angle (h°) value and increase of redness (CIE a*) and chroma (C*) values (p<0.05). BHT, AA, and CE were observed effectively to retard lipid oxidation and increase color stability. BHT and AA revealed significantly (p<0.05) higher thiol content than the control and CE. However, the reduction percentage for thiol content in CE treated patties was lower than the control and AA-treated patties from first to last time of storage. Moreover, inclusion of AA and CE led to significantly (p<0.05) increased heme iron content when compared to BHT and the control. In conclusion, CE can replace the application of AA and BHT while improving lipid stability, heme iron content, and color stableness of fresh beef patties throughout frozen storage.

• Clinical and Experimental Reproductive Medicine
• /
• v.49 no.2
• /
• pp.117-126
• /
• 2022

Objective: This study examined whether the addition of triple antioxidants (3A)-10 µM acetyl-L-carnitine, 10 µM N-acetyl-L-cysteine, and 5 µM α-lipoic acid-in freezing-thawing medium during human sperm cryopreservation using the sucrose vitrification (SuV) and liquid nitrogen vapor (Vapor) techniques could improve post-thaw survival of spermatozoa. Methods: We analyzed 30 samples from healthy human sperm donors. Each sample was allocated into one of five groups: fresh control, SuV, SuV+3A, Vapor, and Vapor+3A. The sperm motility, morphology, viability, intracellular and extracellular reactive oxygen species (ROS) levels, and sperm DNA fragmentation (SDF) were evaluated. Results: The cryopreserved spermatozoa had significantly reduced percentages of motility (p<0.05) and viability (p<0.05). Antioxidant supplementation non-significantly improved these parameters (p>0.05). No significant differences were found in sperm morphology between the fresh and frozen-thawed groups (p>0.05). After freezing, the extracellular ROS levels in the frozen-thawed groups were significantly higher (p<0.05) than in the fresh group. However, we did not find any differences in intracellular ROS parameters among these groups (p>0.05). The SDF was higher in the SuV and Vapor groups than in the fresh group, but without statistical significance (p=0.075 and p=0.077, respectively). Conclusion: Cryopreservation had detrimental effects on sperm motility, viability, and extracellular ROS levels, without changing the morphology or intracellular ROS levels. Antioxidant supplementation was slightly effective in preventing SDF in frozen-thawed spermatozoa.

• Journal of Animal Science and Technology
• /
• v.56 no.3
• /
• pp.8.1-8.6
• /
• 2014

Background: Artificial insemination (AI) can serve as a powerful tool to the sheep owners for making rapid genetic progress of their flock. The AI in sheep is mostly performed using fresh semen with two reasons i) lambing rate following trans-cervical AI with frozen semen is limited by the inability of frozen-thawed sperm to transit the cervix and ii) the need of circumventing the cervical barrier through laparoscope aided intrauterine AI. Therefore, AI with frozen-thawed semen is not as widespread in sheep as it is in other domestic species. However, to get maximum benefits through the use of AI, frozen-thawed semen is a prerequisite because instead of high fertility, the short shelf life of fresh semen coupled with a limitation on the number of insemination doses achievable per unit time restricts the widespread use of individual sires. Therefore, in order to enhance lambing rate, a total of 240 trans-cervical artificial inseminations with frozen-thawed semen were performed in Bharat Merino ewes during autumn season either once in the evening (G-I, 10 h after onset of estrus, n = 100) or twice (G-II, 14 h and 22 h after onset of estrus, n = 140) i.e. once in the morning and again in the evening. Results: The pregnancy rate (proportion of pregnant ewes confirmed by ultrasonography at day 40) and lambing rate (proportion of ewes lambed) were higher in G-II as compared to G-I (26.4 vs 20% and 19.3 vs 10%, respectively). The difference in lambing rates was statistically (P < 0.05) significant. The depth of insemination within cervico-uterine tract had no significant effect on pregnancy and lambing rates. Conclusions: The results indicate that lambing rate in sheep following TCAI with frozen-thawed semen was significantly influenced by time of inseminations. Two inseminations after 14 and 22 h of onset of estrus enhanced the lambing rates of Bharat Merino sheep as compare to single insemination after 10 h of onset of estrus. The TCAI technique with frozen-thawed ram semen is promising and may serve as a valuable tool for genetic improvement of sheep breeds. Research efforts are going on worldwide to overcome the poor fertility following TCAI with frozen-thawed semen.

• Food Science of Animal Resources
• /
• v.40 no.3
• /
• pp.444-460
• /
• 2020

This study was conducted to investigate the effects of freezing and storage temperature (-18℃, -50℃, and -60℃) on the physicochemical properties of pork neck and chicken leg meat in home-scale deep freezers. Pork neck was cut into a thickness of 3 cm (9×9×3 cm, 150 g), individually packed in air-containing packages, and stored at different temperature (-18℃, -50℃, and -60℃) for 6 months. Chicken leg meats were prepared (10 cm long, weighing 70 g) and packed in the same manner. Frozen samples were thawed at 2℃. Physicochemical properties such as thawing loss, cooking loss, water-holding capacity, color, volatile basic nitrogen (VBN), and thiobarbituric acid reactive substances (TBARS) were evaluated. The samples frozen by deep freezing (-60℃) was favorable with respect to thawing loss, color, and VBN. Samples frozen at -60℃ had lower values of thawing loss and VBN than those frozen at -18℃ for all storage periods (p<0.05). Color parameters were more similar to those of fresh meat than to those of samples frozen at -18℃ for 6 months. The TBARS of all samples were below 0.3 mg malondialdehyde/kg, thereby indicating oxidative stability of lipids. Consequently, deep freezing at -60℃ may be acceptable for maintaining the quality of fresh pork neck and chicken leg meat for 6 months without deterioration.