• Title/Summary/Keyword: thawing methods

Search Result 244, Processing Time 0.029 seconds

Freezing Methods of Canine Semen to Achieve Good post-Thaw Viability of Sperm (개 정액의 융해후 정자의 생존율 향상을 위한 동결 방법)

  • 지동범;김용준
    • Journal of Veterinary Clinics
    • /
    • v.17 no.2
    • /
    • pp.420-430
    • /
    • 2000
  • These studies were preformed to investigate the freezing conditions to achieve good post-thaw viability of spend and the practical methods of artificial insemination frozen canine semen. Semen were collected from nine male dogs which had been proved to be fertile in the past and the semen were treated for freezing procedure. Post-thaw motility and viability of canine sperm were evaluated to investigate individual tolerance of freezing, difference among freezing extenders, dif-ference among freezing equipments and freezing conditions, difference between fast and slow cooling rate, difference according to different glycerol concentration, effect of seeding on post-thaw viability, difference according to cutting part of straw, difference according to thawing temperatures, and dif-ference according to media added to thawed semen. Thawed semen for insemination were added with equal volnme of canine capacitation medium (CCM) and the volume of semen and the number per insemination were adjusted as 2-3 ml and $20-30 {\times}10^7,$ respectively. The semen were inseminated in vagina using balloon catheter and en17ryos were cellected from 9 to 11 days after the second Al to d determine fertilization.

  • PDF

Effect of Thawing Rate on the Function of Cryopreserved Canine Sperm (융해 속도가 동결.융해된 개 정자의 기능에 미치는 영향)

  • Kim, Su-Hee;Yu, Do-Hyeon;Kang, Tae-Woon;Kim, Yong-Jun
    • Journal of Veterinary Clinics
    • /
    • v.28 no.6
    • /
    • pp.571-575
    • /
    • 2011
  • Sperm cryopreservation methods have been improved over the last few decades. However, an optimized thawing rate has not yet been established. Therefore, we investigated the effect of thawing rate on sperm function after cryopreservation. The ejaculates collected from beagle dogs were cryopreserved and then thawed at two different thawing rates ($37^{\circ}C$ for 1 min or $70^{\circ}C$ for 15 sec). The thawed sperm were evaluated for motility, viability, morphology, plasma membrane integrity, phosphatidylserine (PS) translocation, and intracellular $H_2O_2$ level. The sperm thawed rapidly at $70^{\circ}C$ showed improved motility, viability, normal morphology, plasma-membrane integrity and non-PS translocation compared to the sperm thawed slowly at $37^{\circ}C$ (P < 0.05). However, the intracellular $H_2O_2$ levels were not significantly different between the rapid- and slow-thawed sperm (P > 0.05). In conclusion, sperm rapid thawing at $70^{\circ}C$ could improve the function of cryopreserved canine sperm, and the appropriate thawing rate would enhance the quality of the cryopreserved sperm.

Evaluation of Physicochemical Deterioration and Lipid Oxidation of Beef Muscle Affected by Freeze-thaw Cycles

  • Rahman, M. H.;Hossain, M. M.;Rahman, S. M. E.;Amin, M. R.;Oh, Deog-Hwan
    • Food Science of Animal Resources
    • /
    • v.35 no.6
    • /
    • pp.772-782
    • /
    • 2015
  • This study was performed to explore the deterioration of physicochemical quality of beef hind limb during frozen storage at −20℃, affected by repeated freeze-thaw cycles. The effects of three successive freeze-thaw cycles on beef hind limb were investigated comparing with unfrozen beef muscle for 80 d by keeping at −20±1℃. The freeze-thaw cycles were subjected to three thawing methods and carried out to select the best one on the basis of deterioration of physicochemical properties of beef. As the number of repeated freeze-thaw cycles increased, drip loss decreased and water holding capacity (WHC) increased (p<0.05) till two cycles and then decreased. Cooking loss increased in cycle one and three but decreased in cycle two. Moreover, drip loss, WHC and cooking loss affected (p<0.05) by thawing methods within the cycles. However, pH value decreased (p<0.05), but peroxide value (p<0.05), free fatty acids value (p<0.05) and TBARS value increased (p<0.05) significantly as the number of repeated freeze-thaw cycles increased. Moreover, significant (p<0.05) interactive effects were found among the thawing methods and repeated cycles. As a result, freeze-thaw cycles affected the physicochemical quality of beef muscle, causing the degradation of its quality.

Studies on cryopreservation of nuclear transplanted mouse embryos (마우스 핵이식란의 동결에 관한 연구)

  • Lee, Byeong-chun;Jo, Choong-ho;Hwang, Woo-suk
    • Korean Journal of Veterinary Research
    • /
    • v.33 no.1
    • /
    • pp.171-178
    • /
    • 1993
  • The present study was carried out to investigate the developmental potency to blastocyst after freezing and thawing of nuclear transplanted 2-cell embryos. The nuclei from 2-, 4- and 8-cell mouse embryos were transferred into enucleated 2-cell embryos, and the reconstituted embryos were submitted to direct current(DC) pulse at output voltage of 2.0 kV/cm for $100{\mu}$ sec to induce cell fusion. The recovery rate and developmental potency to blastocyst after freezing and thawing of nuclear transplanted 2-cell embryos was investigated. 1. The recovery rate of nuclear transplanted 2-cell embryos in normal morphology after freezing and thawing was significantly higher in rapid freezing(DMSO 4.5M) than in slow cooling(p<0.01). 2. When the recovered embryos in normal morphology were cultured in vitro, there were no significant differences in the developmental potency to blastocyst between the freezing methods and the concentrations of cryoprotectant. In summary, these experiments have proved that rapid freezing method(DMSO 4.5M) is effective in nuclear transplanted 2-cell mouse embryos. If improved micromanipulation techniques and freezing are combined, nuclear transplantation technique will contribute to the improvement of productivity in livestock animals.

  • PDF

Effects of Freezing Storage Temperature and Thawing Time on Separation of Leg Meat from Red Snow Crab Chionoecetes japonicus (냉동온도 및 해동시간이 홍게(Chionoecetes japonicus) 다릿살 채육에 미치는 영향)

  • Kim, Byoung-Mok;Jeong, Jee-Hee;Jung, Min-Jeong;Kim, Jong-Chan;Jun, Ki-Hong;Kim, Dong-Soo;Lee, Kwang-Pyo;Jun, Joon-Young;Jeong, In-Hak
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.48 no.5
    • /
    • pp.655-660
    • /
    • 2015
  • This study investigated the effects of freezing storage temperature and thawing time on the separation of leg meat of the red snow crab Chionoecetes japonicus. Crabs were stored at -20, -30, -40, or -50°C for 2 days and thawed for either 5, 10, 20, 30, or 40 seconds. While thawing, there were no significant differences in pH or acidity among the experimental groups, while the volatile basic nitrogen content increased continuously. The redness of samples stored at -20°C was higher than that of the other groups. The overall acceptance of samples stored at -20°C was also the best. These results demonstrate that no-heating methods may be useful for separating red snow crab leg meat.

Frost resistance of porous concrete assuming actual environment (實環境を考慮したポーラスコンクリートの耐凍害性の評価(실제 환경을 고려한 다공질 콘크리트의 내동해성(耐凍害性) 평가))

  • NAKAMURA, Takuro;HORIGUCHI, Takashi;SHIMURA, Kazunori;SUGAWARA, Takashi
    • Proceedings of the Korean Institute of Building Construction Conference
    • /
    • 2008.11a
    • /
    • pp.227-233
    • /
    • 2008
  • Porous concrete has large continuous voids of 20-30 % by volume, and this concrete is attractive as environmental material in Japan i.e. permeable road pavement, river bank protection with vegetation and green roof system which influence thermal environment. It is necessary to confirm the frost resistance when constructing porous concrete structure in cold region. However applicable test method and evaluation criterion of porous concrete has not defined yet. Therefore, the object of this study is to investigate the frost resistance of porous concrete and this investigation attempts to address this concern by comparing 4 kinds of specified freezing and thawing tests methods (JIS A1148 procedure A/B and RILEM CIF/CDF test) in consideration of actual environment. RILEM freeze-thaw tests are different from JIS A1148 freeze-thaw tests, which are widely adopted for evaluating the frost resistance of conventional concrete in Japan, in water absorption, cooling rate, length of freezing and thawing period, and number of freezing and thawing cycles. RILEM CIF test measures internal damage and is primarily applicable for pure frost attack. CDF test is appropriate for freeze-thaw and de-icing salt attack. JIS A1148 procedure A/B showed extremely low frost resistance of porous concrete if the large continuous voids were filled with water and the ice expansion in the large continuous voids set in during cooling. Frost resistance of porous concrete was improved by mixing coarse aggregate (G7) which particle size is smaller and fine aggregate in JIS freezing and thawing tests. RILEM CIF/CDF test showed that freeze-thaw and de-icing resistance of porous concrete was seems to be superior in that of conventional concrete.

  • PDF

A Compound Deterioration Assessment of Concrete Subjected In Freezing-Thawing and Chloride Attack (동결융해와 염해의 복합작용을 받는 콘크리트의 내구성능 저하 평가)

  • 고경택;김도겸;김성욱;조명석;송영철
    • Journal of the Korea Concrete Institute
    • /
    • v.13 no.4
    • /
    • pp.397-405
    • /
    • 2001
  • In clod weather regions, a strong seasonal wind brings sea salts to the land. In addition to it, recently, the spreading amount of deicing salts has increased numerously for purpose of removing snow and ice. Thus the salts environment around concrete structures becomes so severe that various damages of concrete due to applied salts will be brought up. Much of countries such as America, Europe etc. is carried out study for effects of deicing salts on concrete. However, there are not test methods for deterioration of concrete subjected to both freezing-thawing and chloride in Korea. In this study, we carried out test for the compound deterioration subjected to both freezing-thawing and chloride attack, to investigate the effects of sodium chloride on the deterioration of concrete. The test was performed to investigate the effects of cement type, strength and air content on the scaling deterioration of concrete. As a result, the scaling deterioration was accelerated in the presence of salts. And the resistance to scaling was strongly influenced by the type of cement, the strength and air content of concrete.

A Comparative Study on the Extenders for Freezing Canine Semen (개 동결정액을 위한 희석액의 비교연구)

  • Kim Heui-Eun;Lee Jung-won;Kim Nam-Soo;Choi In-Hyuk
    • Journal of Veterinary Clinics
    • /
    • v.10 no.1
    • /
    • pp.11-18
    • /
    • 1993
  • Four extenders such as tris-fructose-citrate, Tris-glucose-citrate, glycine-glucose-citrate and lactose that the more frequently utilized types of semen extenders used for freezing dog semen evaluated with sperm motility, viability and acrosomal score in the processing procedures to prior freezing and after frozen-thawing respectively. Each extender contained 4% glycerol and 20% egg yolk were treated by same methods in dilution, freezing, storage and thawing. The results were obtained as follows; 1. The sperm motility and viability in procedure from dilution to frozen-thawing appeared superiorly with recovery rate of 53.2%, 54.8% in tris-fructose-citrate but appeared inferiorly with recover rate of 8.4%, 8.3% in lactose to others. 2. In the processing procedure course to prior-freezing, glycine-glucose-citrate appeared superiorly with decrease rate of 5.4% in motility, and lactose with decrease rate of 4.6% in viability, but tris-glucose-citrate appeared inferiorly with decrease rate of 12.2%, 11.4% in the sperm motility and viability. 3. During frozen-thawing, tris-fructose-citrate appeared superiorly with decrease rate of 35.2% in motility and 30.7% in viability but lactose appeared inferiorly with decrease rate of 76.7% in motility and 75.7% in viability. 4. The variation of acrosome morphology in the total processing procedures appeared that glycine-glucose citrate were superior with acrosome score of 0.1191$\pm$0.029, that tris-fructose-citrate were inferior with acrosome score of 0.1941$\pm$0.045 to others.

  • PDF

Seminal plasma modulates post-thaw longevity and motility of frozen sperm in dromedary camel

  • Fahimeh Seyedasgari;Behnam Asadi;Ellen Kim
    • Animal Bioscience
    • /
    • v.36 no.12
    • /
    • pp.1821-1830
    • /
    • 2023
  • Objective: This study investigated the effect of adding seminal plasma to frozen-thawed semen on the quality of sperm and pregnancy following insemination in dromedary camels. Methods: In experiment 1, the frozen-thawed semen from 9 collections (3 bulls) was further diluted with either the base extender or homologous seminal plasma (HSP). In the second experiment, a pooled sample of frozen-thawed semen was diluted with either seminal plasma from another three bulls. Live percentage, total and progressive motility, functional and acrosome integrity, and sperm kinematics were evaluated at 15, 60, and 120 minutes post-thawing and compared to the non-treated control. In experiment 3, frozen semen was used to inseminate camels in the following experimental groups: 1-Single insemination with double dose undiluted frozen semen (n = 9); 2-Re-insemination in 6 hours with undiluted semen (n = 13); 3-Single insemination with HSP treated sperm (n = 14). Results: Frozen-thawed sperm diluted in HSP or the non-homologous seminal plasma from Bull C indicated an improvement in all parameters after 1 hour post-thawing incubation (p<0.05). The proportion of total and progressively motile sperm did not drop significantly at 60 minutes post-thawing when diluted with the seminal plasma of Bull C (p>0.05). Double insemination with nontreated sperm and single insemination with HSP-treated sperm resulted in similar pregnancy rates (15.3% vs 21.4%, p>0.05). None of the camels conceived with double-dose single insemination of nontreated sperm. Conclusion: Seminal plasma improves sperm longevity and motility after thawing in dromedary camel with a significant between-bull variation in effect. Low post-thaw sperm longevity might be the cause behind the low pregnancy rates in frozen semen insemination of dromedary camels.

Effects of Different Infusion Frequency of Liquid Nitrogen on Human Embryo Development and Pregnancy Rates after Freezing and Thawing (인간 배아 동결 해빙시 액체질소의 분사속도가 배아 발달 및 임신에 미치는 영향)

  • Kim, Young-Ah;Seo, Seong-Seog;Kim, Mi-Ran;Hwang, Kyung-Joo;Park, Dong-Wook;Jo, Mi-Yeong;Ryu, Hee-Suk
    • Clinical and Experimental Reproductive Medicine
    • /
    • v.28 no.4
    • /
    • pp.287-293
    • /
    • 2001
  • Objective : To investigate the efficacy of high infusion frequency of liquid nitrogen on pregnancy in human embryo after freezing and thawing. Materials and Methods: 150 infertile patients underwent 162 consecutive thawing-ET cycles. In the high infusion frequency group (Group A), 47 patients (50 cycles) underwent cryopreservation with high infusion frequency of liquid nitrogen. In the low infusion frequency group (Group B), 103 patients (112 cycles) underwent cryopreservation with low infusion frequency of liquid nitrogen. We analyzed the clinical characteristics, fertilization rates, development of embryo, good quality embryo ratio, implantation rates, and pregnancy rates between these two groups. Results: There was no difference between the groups with regard to clinical characteristics (mean age, infertility duration, infertility factors, hormone profile), mean number of oocyte retrieval, fertilization rates, and mean embryo number of transfers. The survival rates in group A was 64.9% (228 of 350 embryos), and among the 228 embryos 190 embryos (83.3%) which progressed to the two- to eight-cell stage. After thawing, the embryo numbers were 65 (34.2%), 29 (15.3%), 35 (18.4%), and 37 (19.5%) of grades 1, 2, 3, and above 4, respectively. The survival rates in group B was 63.8% (482 of 755 embryos), and among the 482 embryos 465 embryos (96.5%) which progressed to the two- to eight-cell stage. After thawing, the embryo numbers were 106 (22.8%), 94 (20.2%), 89 (19.1%), and 112 (24.1%) of grades 1, 2, 3, and above 4, respectively. There was no difference in embryo quality change after the freezing-thawing procedure between the groups. Implantation rates (31.1% vs. 34.3%) were not significant. However hCG positive rates in group A (40%) were higher than group B, but not statistically significant. Clinical pregnancy rate (26% vs. 25.9%), on going pregnancy rates (>20 weeks) were not significant (26% vs. 25%). Conclusion: We compared embryo quality change, survival rates, and pregnancy rates between high infusion frequency group and low infusion frequency group and the results were similar between the two groups. Therefore, high infusion frequency of liquid nitrogen for cryopreservation is a worthy method to preserve in human embryos.

  • PDF