• Title, Summary, Keyword: Liquid Semen

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Development of Quantitative Extraction Method of Amygdalin without Enzymatic Hydrolysis from Tonin(Persicae Semen) by High Performance Liquid Chromatography

  • Hwang, Eun-Young;Lee, Sang-Soo;Lee, Je-Hyun;Hong, Seon-Pyo
    • Archives of Pharmacal Research
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    • v.25 no.4
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    • pp.453-456
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    • 2002
  • Tonin(Persicae Semen) is the herb medicine that contains amygdalin as a major ingredient. Amygdalin in water is decomposed into benzaldehyde, HCN, and glucose by emulsin, a hydrolysis enzyme in tonin. A useful and practical method for the optimum extraction condition of amygdalin without enzymatic hydrolysis is required. The extraction yield of amygdalin of natural formula to nin was 0.1 % from crude powders, 1.4% from small pieces, 3.5% from half pieces and 2.4% from whole pieces. The extraction yield of amygdalin of outer shell-eliminated to nin was 0.3% from crude powders, 1.4% from small pieces, and 3.5% from half pieces and whole pieces respectively. The extraction yield of amygdalin was most high when using the size larger than half.

Prevention of Epimerization and Quantitative Determination of Amygdalin in Armeniacae Semen with Schizandrae Fructus Solution

  • Joo, Woo-Sang;Jeong, Ji-Seon;Kim, Hyo-Geun;Lee, Yong-Moon;Lee, Je-Hyun;Hong, Seon-Pyo
    • Archives of Pharmacal Research
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    • v.29 no.12
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    • pp.1096-1101
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    • 2006
  • Armeniacae Semen not only contains amygdalin, but emulsin also, which is an enzyme that hydrolyzes amygdalin. The extraction yield of amygdalin from Armeniacae Semen was low, due to the presence of emulsin, when extracted with water. When Schizandrae Fructus solution was used as the extractant; however, amygdalin was almost completely extracted, regardless of the cutting size, due to the absence of the influence of emulsin. In addition, when the crude powder or small piece forms were used with Schizandrae Fructus solution, on epimerization of the D-amygdalin into neoamygdalin occurred. D-amygdalin and its conversion product, neoamygdalin, were quantitatively analyzed by reverse-phase, high-performance liquid chromatography (HPLC), with an optimized eluent of 10 mM sodium phosphate buffer (pH 2.3), containing 11.5% acetonitrile. The concentration and detector response were linearly correlated over the range 0.05 to 2 mM. The detection limits for both D-amygdalin and neoamygdalin were approximately $5\;{\mu}M$ for the amount injected.

Production of Pups Following Artificial Insemination by Canine Intrauterine Inseminator (개 자궁내 인공수정기에 의한 인공수정 후 산자생산)

  • 공일근;조성균;임용택;이상인;위성하
    • Journal of Veterinary Clinics
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    • v.16 no.2
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    • pp.375-380
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    • 1999
  • This study was conducted to develop an intrauterine inseminator (IUI) to deposit of frozen semen into uterus and to evaluate the results obtained after artificial insemination by IUI. Two Japanese spitzs (2 to 4 years of age) were used as semen donors. Semen was collected by manual masturbation into sterile glass collection tubes and separated into 3 fractions with only the sperm-rich fractions retained for further examination. Sperm motility >70%, sperm concentration of 200 to $400{\times}10^6 cells/ml$$\times$g for 5 min and poured out the suspended solution, and then diluted with 2 ml Tris-buffer which was consisted of 2.4 g Tris, 1.4 g citric acid, 0.8 g glucose, 0.1 $\mu\textrm{g}$/ml streptomycin, 100 IU/ml penicillin, 20 ml egg yolk to 100 ml mili-Q water (Ext I) or supplemented with 8 ml glycerol and 1 ml Equex STM paste to 100 rnl (Ext II). The diluted semen was cooled to 5$^{\circ}C$ in cold room, where the temperature in the sample reached 5$^{\circ}C$. Two h after beginning the cooling procedure, 2 ml of Ext II, also at 5$^{\circ}C$, was added and mixed by gently reversing the tubes several times during 1 h. The final sperm concentration for freezing was approximately $50{\times}10^6 cells/ml$. After equilibration, the semen was loaded into 0.5 ml straw and frozen on the liquid nitrogen vapour in styrofoam box. The straws were thawed at 7$0^{\circ}C$ for precisely 6 sec. After thawing of each straw, the frozen semen can survived over 50% motility. All the females were inseminated twice with 1 ml of $25{\times}10^6 cells/ml$

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Effects of Kinds of Cryoprotectants on the Characteristics of Frozen Fowl Semen (닭 정액 동결 시 동결 보호제가 정액 성상에 미치는 영향)

  • Choi, Jin Seok;Shin, Dan-Bi;Ko, Yeoung-Gyu;Do, Yoon-Jung;Byun, Mijeong;Park, Soo-Bong;Seong, Hwan-Hoo;Kim, Hyun;Kong, Il-Keun;Kim, Sung Woo
    • Korean Journal of Poultry Science
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    • v.40 no.3
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    • pp.171-178
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    • 2013
  • The purpose of this study was to evaluate the sperm viability, normal acrosome and mitochondrial activity in the frozen-thawed fowl semen by different cryoprotectants. The experiment was carried out on 10 sexually adult roosters of Ogye. The semen was collected twice a week and pooled semen was diluted 1:1 EK extender containing no cryoprotectant at $5^{\circ}C$. After equilibration for 30 minutes, diluted chicken semen was diluted 1:1 extender containing either 7% dimethylacetamide (DMA), 7% dimethylformamide (DMF) or 7.5% methylacetamide (MA) at final concentration and was put in 0.5 mL plastic straws and frozen for 30 minutes by exposure to liquid nitrogen vapor 4 cm above the surface of liquid nitrogen, followed by plunging into liquid nitrogen. Frozen semen was thawed in water bath at $5^{\circ}C$ for 2 minutes. For cytometric analysis, the frozen-thawed semen was diluted with EK extender to a final concentration of 90 million spermatozoa per mL. Sperm membrane integrity was evaluated as SYBR-14 and propidium iodide (PI). Acrosome integrity was assessed with fluorescein isothiocyanate-labeled PSA and PI. The percentage of mitochondrial function was estimated by using Rhodamine123 (R123) and PI. In conclusion, freezing rooster semen by using 7% DMF as cryoprotectant was significantly highest in rates of survival and mitochondrial function while its rate of damage of acrosome was significantly lowest. As a result, DMF is the cryoprotectant that has the lowest influences on sperm membranes and acrosome integrity. Therefore it could be used for freezing method of animal genetic conservation method for poultry diversity.

Rhei Rhizoma Mainly Blended Prescriptions According to the Fomula, Manipulation, Related Co-herb in Dongeuibogam (『동의보감(東醫寶鑑)』 중 대황(大黃)이 주약(主藥)으로 배오(配伍)된 방제(方劑)의 제형(劑形), 포제(?製), 약대구성(藥對構成)에 따른 활용(活用))

  • Joh, Hae-In;Kook, Yoon-Bum
    • Herbal Formula Science
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    • v.25 no.4
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    • pp.553-574
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    • 2017
  • The purpose of this study is to find out effects of prescriptions according to the formula, manipulation of Rhei Rhizoma, configuration. The following results were reached through investigations on the prescriptions using Rhei Rhizoma as a main component. Objectives : Analysis of prescriptions According to the formula : Liquid Extract Prescriptions were used widely on three parts to treat fever and damp heat in the interior organs. Powder Prescriptions were taken with hot water, thin porridge, tea etc. to treat damp heat, congestion of phlegm, acute episodes. Liquid Mixed Pill treat congestion of QI, damp heat, phlegm. Honey Mixed Pill treat accumulated fever, distension, acute excessive fever. Paste Pill treat blood stagnation, excessive toxic-fever, epidemic diseases. External Application treat inflammation by injury, swelling due to severe fever by internal damage. Methods : Analysis of prescriptions According to the manipulation of Rhei Rhizoma : Prescriptions including Liquor processed Rhei Rhizoma treat excessive toxic-fever, congestion of phlegm, blocking orifices on the upper side. Steamed Rhei Rhizoma strengthen effects of making evacuate and cooling of heat. Processed Rhei Rhizoma with vinegar strengthen effects of removing blood stagnation by activating blood movement, releasing gathering. Results : Analysis of prescriptions According to the Composition of Rhei Rhizoma : 41% of the total prescriptions were on the area of less than 20%. In case of lower groups show increased frequency of combination with Pharbitidis Semen, Persicae Semen, Scutellariae Radix and manipulation of baking, steaming, roasting. In case of higher groups show increased frequency of treating excess syndrome, critical illness, acute severe illness, and using proccesed Rhei Rhizoma with vinegar. Treatment of damp heat on the liver and gallbladder, disorder of the spleen and stomach is done mostly by prescriptions on the area of less than 30%. Conclusions : Rhei Rhizoma-Coptidis Rhizoma pair treat damp heat, heat toxins in blood, and Constipation caused by excessive heat. Rhei Rhizoma-Glycyrrhizae Radix pair relieve effects of Rhei Rhizoma passing blocked feces, removing the poison, activating blood movement, releasing gathering with the effects of Glycyrrhizae Radix relaxing tension by harmonizing Middle. Rhei Rhizoma-Magnoliae Cortex pair are used to treat damp heat in middle area, excessive heat in the stomach and intestine. Rhei Rhizoma-Pharbitidis Semen pair act on both blood system and QI system treating edema, damp, stagnation, heat toxins, feces. Rhei Rhizoma-Persicae Semen pair treat blood stagnation with fever on blood system.

Establishment of Cryopreservation of Leopard Cat Semen Collected by Electro-ejaculation Method

  • Ha, A-Na;Jo, A-Ra;Kim, Yu-Gon;Yoon, Jin-Ho;Bang, Jae-Il;Deb, Gautam K.;Fakruzzaman, M.;Lim, Yang-Mook;Yong, Hwan-Yul;Kong, Il-Keun
    • Journal of Embryo Transfer
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    • v.26 no.4
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    • pp.245-250
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    • 2011
  • The aim of this study was to evaluate the post-thawed characteristics of leopard cat semen. In this experiment, semen was collected from two leopard cats (A and B) at wild animal center in Seoul Grand Park in Korea. After collection, the sperms were washed with D-PBS and diluted by the freezing medium (Irvine science, USA) and stored in liquid nitrogen. The post-thawed concentration was $357{\times}10^6sperms/ml$ for A and $97{\times}10^6sperms/ml$ for B. The viability of post-thawed sperm from A and B individual was 24.0% and 19.0%, respectively. Pre-freezing motility of A and B individual semen was 68.54% and 56.65. Leopard cat A had more normal sperm than that of B (69.5% vs. 54.5%). Acrosome integrity analysis detected live (14.5% vs. 9.0%), damage (39.0% vs. 44.0%) and dead (46.0% vs. 47.0%) in leopard cat A and B, respectively. The present results concluded that leopard cat semen can be collected successfully by electro-ejaculation method and cryopreserved successfullyfor future use in different assisted reproductive technologies. The cryopreservation protocol needs to be modified for increasing post-thawed viability of leopard cat spermatozoa.

Characterization of Bacteria and Their Antibiotic Sensitivities in Porcine Liquid Semen (돼지 액상정액 내 세균오염과 항생제 감수성에 관한 연구)

  • Ryu, Jae-Weon;Cho, Kyu-Ho;Hong, Joon-Ki;Kim, Myung-Jick;Park, Jun-Chul;Jung, Il-Byung;Kim, In-Cheul
    • Journal of Animal Science and Technology
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    • v.50 no.6
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    • pp.793-798
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    • 2008
  • The semen collection process in the porcine is far from being a sterile procedure. Consequently, porcine ejaculates commonly contain bacterial contaminants. The aim of this study is to identify the bacteria in porcine semen and to find the antibiotics resistance of bacteria. Twelve porcine originating from four AI center were used to collect semen. Bacteria were identified by automated instrument for rapid organism identification system and bacterial sensitivities of 8 antibiotics were tested. The Bacterial contaminants of Staphylococcus genus(37.8%), Proteus genus(7.0%), Bacillus genus (6.1%), Pasteulla genus(5.7%), Acinetobacte genus(5.2%), Serratia genus(4.3%) and others(33.9%) were frequently isolated. However, amikacin showed higher antibiotic sensitivity than other antibiotics. General sanitation protocols can contribute partly to inhibit the bacterial contamination, with monitoring boar housing, semen collection areas and the extended semen. But, proper selection of preservative antibiotics by microbial sensitivities can minimize the influence of bacteria.

Effects of Seeding during Freezing Procedure on Post-Thaw Viability and Acrosome Integrity of Boar Spermatozoa (돼지정액 동결중 식빙처리가 융해후 정자생존율 및 침체형태에 미치는 영향)

  • Kim Yong-jun;Kim Yong-hwan;Lee Young-jun;Kim Sue-hee;Ji Dong-beom
    • Journal of Veterinary Clinics
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    • v.21 no.4
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    • pp.363-368
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    • 2004
  • To investigate the effects of seeding during freezing procedure on post-thaw viability, motility, and acrosome integrity of boar spermatozoa, semen from 5 Yorkshire boars were collected for this experiment. Raw semen were diluted with Merck I, subsequently added with cooling diluent containing lactose and egg yolk and with freezing diluent containing glycerol. The diluted semen were frozen on the rack in the styrofoam box filled with liquid nitrogen at the distance of 5 cm or I cm above LN2 level. Seeding was performed to only a group of straws frozen at 5 cm away on the surface of LN2. The frozen semen were thawed in $50^{\circ}C$C water and the viability and local motility were analyzed by sperm analysis imaging system. A part of thawed semen was taken for the examination of morphology of apical ridge of the acrosome to compare with the effect of seeding between the seeding-treated and non treated groups. I. Post-thaw viability was considerably higher in seeding-treated sperm than non-seeding group (p<0.01), however, no difference of local motility was obtained among the groups. 2. At three hours after thawing, viability was also higher in seeding-treated group than non-treated group (p<0.05), along with no difference of motility among the groups. 3. Higher normal acrosome integrity was obtained in the seeding-treated sperm than non-treated groups (p<0.01). 4. Between non-seeded groups, higher normal acrosome integrity was obtained in the sperm group frozen at 5cm upper on the surface of LN2 than that frozen at 1cm away (p<0.01). These results indicated that seeding treatment during freezing boar spermatozoa was beneficial to post-thaw viability and normal acrosome integrity.

닭 정액의 보존온도 및 희석배율이 수정률에 미치는 영향

  • 김학규;나재천;최철환;장병귀;상병돈;이상진
    • Proceedings of the Korea Society of Poultry Science Conference
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    • pp.107-108
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    • 2003
  • This study was conducted to investigate the effects of dilution rate and stored temperature of semen at 5, 25 and 35$^{\circ}C$ on fertility in liquid rooster semen. At 5$^{\circ}C$ cold temperature, no significant difference were found in sperm mobilities on dilution rate(1:1, 1:3, 1.6) among treatments. Sperm mobility for the conservation of 3 hours at 25∼35$^{\circ}C$ were significantly higher for 1:3 and 1:6 dilution rate(semen:diluent) groups than for 1:1 dilution rate group(P<0.05). In Fertility results after artificial insemination with the conservation of 3 hours at 5∼25$^{\circ}C$ temperature, no significant difference were found in fertility on dilution rate among treatments. Fertilities after artificial insemination with the conservation of 3 hours at 35$^{\circ}C$ were significantly higher for 1.3 and 1:6 dilution rate(semen:diluent) groups than for 1:1 dilution rate group(P<0.05).

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Effect of BTS and Androhep during Storage Times on the Kinematics and Capacitation Status in Liquid Boar Semen (BTS와 Androhep이 보존 기간 동안 액상 정액의 운동역학 및 수정능 획득에 미치는 영향)

  • Kim, Yun-Hee;Park, Yoo-Jin;Yoon, Sung-Jae;Kwon, Woo-Sung;Kim, Sang-Hyun;Pang, Myung-Geol
    • Reproductive and Developmental Biology
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    • v.34 no.3
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    • pp.241-246
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    • 2010
  • The objective of this study was to investigate the effect of storage time on fresh boar semen in Androhep and Beltsville Thawing Solution (BTS). Boar semen samples extended in each extender were stored at $17^{\circ}C$ up to 4 days. Sperm motility kinematics was evaluated by computer assisted sperm analyzer (CASA) and capacitation status by chlortetracycline (CTC)/Hoechst 33258 staining. Sperm motility (%) was not decreased during storage in BTS and Androhep. No significant difference between extenders was observed. Only significant differences in kinematic parameters on linearity during storage were found. The percentage of dead sperm significantly decreased during storage (p<0.05). Also the percentage of noncapacitated, capacitated, and acrosome-reacted sperm significantly modified during storage (p<0.05). However, there was no significant difference between extenders except proportion of capacitated sperm. This finding supported that modification in these parameters was not significantly different between extenders during this short-term storage. Our finding strongly indicated that both Androhep and BIS maintained favorable conditions for motility, motility kinematics, and capacitation status during short-term storage. Despite modifications in some parameters were apparent during sperm storage in extenders, these may not affect the fertilizing capacity of boar semen.