• Title/Summary/Keyword: Liquid Semen

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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.

Seminal Attributes and Semen Cryo-banking of Nepalese Indigenous Achhami (Bos indicus) Bull under Ex-situ Conservation

  • Jha, Pankaj Kumar;Sapkota, Saroj;Gorkhali, Neena Amatya;Pokharel, Bhoj Raj;Jha, Ajeet Kumar;Bhandari, Shishir;Shrestha, Bhola Shankar
    • Journal of Animal Reproduction and Biotechnology
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    • v.34 no.4
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    • pp.272-279
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    • 2019
  • The study was conducted to evaluate the seminal attributes and cryobanking of Achhami (Bos indicus) bull semen. Of two Achhami bulls, 8 ejaculates from each bull were evaluated for seminal attributes. For semen freezing and cryo-banking, 4 ejaculates (having ≥2 mL semen volume, ≥75% of sperm motility and ≥1,000 × 106 cells/mL of sperm concentration) from each bull were used. Semen samples were diluted in egg-yolk-tris-citrate extender using a two-step dilution protocol, and were frozen in liquid nitrogen (LN2) vapour in a styrofoam box. The mean semen volume, colour, sperm mass activity, motility, viability, concentration, abnormal acrosome, midpiece and tail and, abnormal head of two Achhami bulls were 4.4 ± 0.5 mL vs. 2.5 ± 0.2 mL, 2.5 ± 0.1 vs. 2.4 ± 0.1, 3.5 ± 0.1 vs. 3.5 ± 0.1, 77.0 ± 1.1% vs. 78.3 ± 1.3%, 94.4 ± 0.5% vs. 91.0 ± 0.6%, 1137.7 ± 73.7 × 106 cells/mL vs. 1060.0 ± 44.3 × 106 cells/mL, 10.2 ± 0.5% vs. 10.3 ± 0.5% and 6.7 ± 0.5% vs. 8.2 ± 0.3%, respectively. The post-thawed sperm motility and viability were 53.0 ± 2.0% vs. 50.0 ± 0.0% and 80.2 ± 0.4% vs. 73.2 ± 0.7%, while evaluating by computer-assisted sperm analysis (CASA) system, the percentage of the progressive motility, fast motility, slow motility, local motility and immotile sperm were 75%, 68%, 7.4%, 16.6% and 8.6%, respectively. A total number of 620 doses semen straw were cryo-banked. Due to the acceptable post-thawed sperm motility and viability recorded, cryopreservation of Achhami semen is hereby recommended so as to preserve the Achhami breed. For further validation, the fertility will be observed from the produced frozen semen.

In-Vitro Fertilization and Culture of Pig Oocytes Matured In-Vitro by Liquid Boar Sperm Stored at 4$^{\circ}C$

  • Kim, M. Y.;Y. J. Yi;Y. J. Chang;Park, C. S.
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.63-63
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    • 2003
  • This study was carried out to investigate the effects of liquid boar sperm stored at 4$^{\circ}C$ on sperm motility, normal acrosome, and in-vitro fertilization and culture of pig oocytes matured in-vitro. The sperm-rich fraction (30~60 ml) of ejaculate was collected into an insulated vacuum bottle. Semen was slowly cooled to room temperature (20~23$^{\circ}C$) by 2 h after collection. Semen was transferred into 15 ml tubes, centrifuged at room temperature for 10 min at 800$\times$g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5 ml of lactose, egg yolk and N-acetyl-D-glucosamine (LEN) diluent to provide 1.0$\times$10$^{9}$ sperm/ml at room temperature. The resuspended semen was cooled in a refrigerator to 4$^{\circ}C$ and preserved for 5 days to examine sperm motility and normal acrosome. The medium used for oocyte maturation was modified tissue culture medium (TCM) 199. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. Oocytes were inseminated with liquid boar sperm stored at 4$^{\circ}C$ for 2 days after collection. Oocytes were coincubated for 6 h in 500 ${mu}ell$ mTBM fertilization media with 0.2, 1, 5 and 10$\times$10$^{6}$ /ml sperm concentration, respectively. At 6 h after IVF, oocytes were transferred into 500 ${mu}ell$ Hepes-buffered NCSU-23 culture medium for further culture of 6, 48 and 144 h. There were significant differences in sperm motility and normal acrosome among preservation days and incubation times, respectively. The rates of sperm penetration and polyspermy were higher in 5 and 10$\times$10$^{6}$ sperm/ml than in 0.2 and 1$\times$10$^{6}$ sperm/ml. Male pronuclear formation was lower in 0.2$\times$10$^{6}$ sperm/ml than in 1, 5 and 10$\times$10$^{6}$ sperm/ml. Mean numbers of sperm in penetrated oocyte were highest in 10$\times$10$^{6}$ sperm/ml compared with other sperm concentrations. The rate of blastocysts from the cleaved oocytes (2~4 cell stage) was highest in 1$\times$10$^{6}$ sperm/ml compared with other sperm concentrations. In conclusion, we found out that liquid boar sperm stored at 4$^{\circ}C$ could be used for in-vitro fertilization of pig oocytes matured in-vitro. Also, we recommend 1$\times$10$^{6}$ ml sperm concentration for in-vitro fertilization of pig oocytes.

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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.

Effects of Glycerol Concentration on Viability of Frozen-thawed Canine Spermatozoa

  • Shin, Young-Jee;Son, Jung-Min;Lim, Young-Hwan;Kim, Young-Sil;Lee, Doo-Soo;Yoon, Ki-Young;Shin, Sang-Tae;Cho, Jong-Ki
    • Journal of Embryo Transfer
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    • v.23 no.2
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    • pp.115-118
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    • 2008
  • Glycerol is the cryoprotectant most frequently used to freeze semen in several of species. The objective of the present study was to compare the effect of three different glycerol concentrations (4, 6 or 8%, v/v) on frozen-thawed dog sperm survival rate. Ejaculates from 9 dogs collected by digital manipulation were pooled and assessed by macroscopic and microscopic criteria. Semen was divided into 3 aliquots, which were centrifuged and the sperm pellets rediluted with first Tris-glucose-citric acid extender. After 1 h cooling at $4^{\circ}C$, second extender containing 4, 6 or 8% glycerol was added, respectively. The semen was loaded into 0.25 ml straws and frozen and stored in liquid nitrogen and thawed. Sperm vigor, live:dead spermatozoa ratio using HOS test, and sperm morphology using $Spermac^{(R)}$ stain were evaluated. After thawing, there were no significant differences among groups in vigor, viability and morphology. In conclusion, the three glycerol concentrations (4, 6 or 8%) can be used successfully in cryopreservation of canine semen. Therefore the use of 4% glycerol in the extender has less toxic effect and reduces of freezing injuries.

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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    • 2003.11a
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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 green tea extract in extender of Simmental bull semen on pregnancy rate of recipients

  • Susilowati, Suherni;Sardjito, Trilas;Mustofa, Imam;Widodo, Oky Setio;Kurnijasanti, Rochmah
    • Animal Bioscience
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    • v.34 no.2
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    • pp.198-204
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    • 2021
  • Objective: The aim of this study was to ascertain the effects of adding green tea extract (GTE) to skim milk-egg yolk (SM-EY) extender on both the quality of post-thawed bull semen and the pregnancy rates of the recipient cows. Methods: Twelve ejaculates from four Simmental bulls, aged 3 to 5 years and weighing 900 to 950 kg, were diluted SM-EY extender, added with 0, 0.05, 0.1, and 0.15 mg GTE/100 mL extender and then frozen. After four weeks storage in liquid nitrogen, the sperm were thawed and evaluated for viability, motility, intact plasma membrane (IPM), and DNA fragmentation. Meanwhile, the estrus cycles of 48 recipient cows were synchronized by intramuscular administration of a single injection of 5 mg prostaglandin F2α. Estrus cows were divided into four equal groups and inseminated artificially 18 to 20 h after the onset of estrus by using semen from each extender group. Pregnancy was diagnosed by measuring serum progesterone levels at 21 days, followed by transrectal palpation 90 days after insemination. Results: The findings revealed that adding 0.1 mg of GTE/100 mL extender produced the highest percentages of sperm viability (70.67%±1.75%), motility (69.17%±1.47%), and IPM (69.23%±1.21%) and the lowest percentage of DNA fragmentation (3.00%±0.50%). The pregnancy diagnosis revealed that all cows (36/36) inseminated using frozen semen in GTE addition extender were pregnant (pregnancy rate 100%), whereas the pregnancy rate of the control group was 83.33% (10/12). Conclusion: It may be concluded that 0.1 mg GTE/100 mL extender yields the best quality of spermatozoa and that all variants doses of GTE in extender produce a higher pregnancy rate among recipient cows.

Study on the Preservation of Liquid Boar Semen with $\textrm{BF}_5$ and Butschwiler Diluents (희석액 $\textrm{BF}_5$ 엔오투와 Butschwiler를 이용한 돼지 액상정액 보존에 관한 연구)

  • 천용민;박창식;서길웅;이규승
    • Journal of Embryo Transfer
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    • v.11 no.2
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    • pp.159-166
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    • 1996
  • 본 연구는 돼지 액상정액을 인공수정용 100ml 플라스틱 병에 보존하면서 BF5희석액과 Butschwiler 희석액 간에 보존 온도별 차이를 조사하고, BF5 희석액에서의 글리세롤 농도의 효과를 조사하여 돼지 액상정액을 좀더 장기간 사용할 수 있는 방법을 찾고자 실시하였다. 돼지 액상정액을 5$^{\circ}C$ 냉장고에 보존하면서 조사한 바에 의하면, 37$^{\circ}C$에서 0.5 및 2시간 배양후의 정자운동성은 전체 보존기간동안 BF5 희석액이 Butschwiler 희석액보다 유의하게 (P<0.05) 높게 나타났고, 정상첨체비율은 두 희석액간에 차이가 없었다. 돼지 액상정액을 15$^{\circ}C$에 보존하면서 조사한 바에 의하면, 3일부터 7일 보존시 까지 정자운동성과 정상첨체비율에 있어서 Butschwiler 희석액이 BF5 희석액보다 유의하데 높게 나타났다. BF5 희석액을 이용한 돼지 액상정액의 글리세롤 농도의 효과에 있어서는 최종 글리세롤 농도가 0, 2, 3, 및 5% 보다 1%일 때 가장 높은 정자운동성과 정상첨체비율을 나타내었다. 분만율, 복당 생존자돈수 그리고 출생시 평균 생시체중은 BF5 희석애과 Butschwiler 희석액간에 차이가 없었다. 이상의 연구 결과를 종합해 볼 때 BF5 희석액을 5$^{\circ}C$에서 Butschwiler 희석액은 15$^{\circ}C$에서 6-7일 동안 돼지 액상정액을 보존할 수 있었다.

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Effect of Cholesterol and Serum Albumin on Sperm Ability and Lipid Peroxidation during the Storage of Miniature Pig Sperm (미니돼지정액의 보존 시 콜레스테롤과 혈청 알부민이 정자 성상과 지질 과산화에 미치는 영향)

  • Kim, Dong-Woo;Lee, Yong-Seung;Yoo, Han-Jun;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Journal of Embryo Transfer
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    • v.26 no.1
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    • pp.71-78
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    • 2011
  • This study was undertaken to find out the effect of cholesterol and serum albumin on sperm ability and lipid peroxidation levels period to the liquid storage of miniature pig sperm. Ejaculated semen from miniature pigs was collected by gloved-hand method into a pre-warmed ($37^{\circ}C$) thermos bottle, and extended with Modena solution {with and without BSA, methyl-beta-cyclodextrin (-cholesterol) and cholesterol loaded cyclodextrin (+cholesterol)}. Each semen was assessed for viability (SYBR-14/PI staining) and acrosome intactness, intensity and capacitation status by chlorotetracycline (CTC) staining at 1, 3, 5, 7 and 10 days of storage. At for the effects of cholesterol and serum albumin on lipid peroxidation, semen were incubated with $H_2O_2$ ($10\;{\mu}M$), and lipid peroxidation level were measured by flow cytometry using the lipid peroxidation reporter probe $C_{11}-BODIPY^{581/591}$. The result, lipid peroxidation level in sperm added with cholesterol were lower in $10\;{\mu}M$ $H_2O_2$ compared to the added sperm with serum albumin. Also, added cholesterol to sperm had significant (p<0.05) higher viability when storage for 7 and 10 days and lower when 10 days of storage percentage of acrosome-reacted sperm (AR pattern) in acrosome state as say result compared to other treated groups. In conclusion, role of cholesterol during lipid storage in miniature pig spermatozoa was protected boar spermatozoa from lipid peroxidation prior to lipid storage. Addition serum albumin during lipid storage in sperm may be induce sperm membrane damage by lipid peroxidation. Therefore, addition of cholesterol to miniature pig sperm will be lead to extension of liquid storage periods.

Current Status of Swine Artificial Insemination(AI) in Korea (한국의 돼지 인공수정 현황)

  • Kim, In-Cheul;Sa, Soo-Jin;Kang, Kwon;Kim, Sang-Hyun;Bae, Sang-Jong;Kim, Dae-Sil;Kim, Si-Joo;Min, Chen-Sik;Son, Jung-Ho;Chung, Ki-Hwa
    • Reproductive and Developmental Biology
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    • v.35 no.3
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    • pp.227-232
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    • 2011
  • This survey was conducted to investigate the current status of swine artificial insemination(AI) centers registered as 'semen processing business' in Korea. The survey responses were collected by direct visitation or telephone conversation for 5 months from May through September in 2008. The survey showed that sixty-four AI centers were enrolled in local government and those of fifty-two AI centers were under operation. Forty-nine AI centers surveyed owned a total of 3,334 boars and the Duroc breed accounted for the highest rate(73.1%) of all boar breeds. In type of ownership, agricultural management corporations was the highest(42.3%) and followed by private ownership(34.6%). Large-scale AI centers in terms of own over 151 boar were surveyed as 5.9% and most AI centers own less than 100 boars(86.5%). The average number of boars per AI center was 68. The amount of liquid semen provided by 52 AI centers were 1,791,000 doses and each AI center provides average of 39,000 does, which is represented for 90% consumption by sows in Korea.