• Title, Summary, Keyword: Boar

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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.

Adaptation of the Hypoosmotic Swelling Test to Evaluate Membrane Integrity of Boar Spermatozoa

  • Jang, Hyun-Yong;Cheong, Hee-Tae;Hwang, Hwan-Sub;Kim, Jong-Taek;Park, Choon-Keun;Lee, Hak-Kyu;Yang, Boo-Keun
    • Reproductive and Developmental Biology
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    • v.31 no.2
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    • pp.121-126
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    • 2007
  • The objective of this study was to establish the optimal conditions for hypoosmotic swelling (HOS) test to assess the functional integrity of the membranes of boar fresh or frozen/thawed spermatozoa. When pooled semen sample was incubated for 30 min at $37^{\circ}C$ with different test solution of varied osmolarity, the highest percentage of HOS positive spermatozoa was observed in a 150 mOsmol fructose/Na-citrate solution (33.6%). Incubation time did not affect significantly the score of HOS positive spermatozoa observed in a 150 mOsmol fructose/Na-citrate solution at $37^{\circ}C$, but the osmolarity affected the score of HOS positive spermatozoa under the same condition above. Fresh semen was significantly better than frozen/thawed semen in semen parameters evaluated such as motility, viability, membrane integrity and lipid peroxidation (p<005). In the relationships of sperm parameters, motility vs viability, motility vs membrane integrity and viability vs membrane integrity were positively correlated ($0.82{\sim}0.94$) but lipid peroxidation vs other estimated factors was negatively correlated ($- 0.90{\sim}- 0.98$). Among the evaluation methods, motility vs Viability, motility vs membrane integrity and lipid peroxidation vs other estimated factors were significantly correlated (p<0.05). These results of this. study indicate that the optimal condition of HOST in boar spermatozoa is a 150 mOsmol fructose/Na-citrate solution for 30 min incubation at $37^{\circ}C$ and HOST can substitute the examination of motility, viability and lipid peroxidation.

A survey of viruses associated with reproductive failure in boar semen in Korean artificial insemination centers (국내 인공수정센터의 웅돈에 대한 번식 관련 바이러스 조사)

  • Kim, Yeong-Hun;Chun, Bong-Su;Kim, Sung-Jae;Han, Jeong-Hee
    • Korean Journal of Veterinary Service
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    • v.34 no.2
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    • pp.111-116
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    • 2011
  • Artificial insemination (AI) of swine is a very useful reproductive tool and that offers convenience in the Korean swine industry. Since many viruses have been reported to be excreted through boar semen, we investigated the presence of antibodies and antigens against viruses causing reproductive failure in semen of boar in 349 semen samples collected from six Korean AI centers. Viral antigens were detected by polymerase chain reaction (PCR) or reverse transcription-PCR predominantly. The results was as follows. The major reproductive failure causing factor was porcine circovirus type 2 (PCV2), followed by porcine reproductive and respiratory syndrome virus (PRRSV) ($X^2$=166.64, P<0.001). PCV2 and PRRSV, Japanese encephalitis virus (JEV), encephalomyocarditis virus (EMCV) was detected in 73 samples (20.9%), 44 samples (12.6%), 4 samples (1.1%), 3 samples (0.9%), respectively and porcine parvovirus in one sample (0.3%) Classical swine fever virus (CSFV), bovine viral diarrhea virus and Aujeszky's disease virus (ADV) were not detected. Enzyme-linked immunosorbent assay was carried out in 111 serum samples from three AI centers. In most pigs, antibodies response was showed prominently in CSFV (105 sera, 94.6%) ($X^2$=82.580, P<0.001), followed by, in PRRSV (100 sera, 90.1%), PCV2 (92 sera, 90.1%), and PPV (8 sera, 82.9%). ADV antibody was not detected. Thus, the experimental results will be used for the base data, with respect to the state of viral stillbirth in general pig farms, as well as AI centers and breeding farms in Korea.

Effect of Monothioglycerol on ROS Inhibition, Mitochondrial Activity, and DNA Integrity in Frozen-thawed Miniature Pig Sperm (Monothiolglycerol이 동결 융해 후 미니돼지 정자의 활성산소 억제, 미토콘드리아 활성 그리고 DNA Integrity에 미치는 영향)

  • Park, Soo-Jung;Kim, Dae-Young
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.265-271
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    • 2013
  • Cryopreservation and in vitro fertilization (IVF) protocols are important in genetic studies and applications to transgenic animals. Various studies about boar sperm cryopreservation have been studied for a long time. Those were about the use of extenders, the choice of sugars, the cooling and warming rates. The factors that influence the boar sperm are the dramatic changes in temperatures, osmotic and toxic stresses, and reactive oxygen species (ROS) generation. Among these factors, ROS generation is the main damage to DNA which is a principal genetic material and the most important for the practical applications. So we wondered whether ROS generation could be reduced. In previous study, monothioglycerol (MTG) was essential for the culture of embryo stem cells. Therefore we added MTG in the freezing extender based on lactose-egg yolk (LEY) with trehalose. For the assessment of the frozen-thawed sperm, we focused onmotility, membrane integrity and DNA damage. First, we used a computer-aided sperm analysis system for overall conditions of sperm such as motility and viability. Then we performed the sperm chromatin structure assay for DNA integrity and hypo-osmotic swelling test for membrane integrity. And our result showed the existence of MTG in the freezing extender caused less damage to DNA and higher motility in frozen-thawed boar sperm. Also we checked a relative antioxidant activity of MTG in modified Modena B extender. We concluded that this reagent can activate sperm mitochondria at MTG $0.2{\mu}M$, contribute to sperm motility and DNA integrity but there was no significant difference on membrane integrity. Also antioxidant activity of MTG in modified Modena B extender was proved.

Effect of dietary isoflavones of soybean by-product on estrogen and testosterone levels in mouse

  • Park, Sungkwon;Choi, Hojun;Seo, Jinyoun;Cho, Sangrae;Kim, Jungsang;Hong, Sung Wook;Park, Changseok
    • Korean Journal of Agricultural Science
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    • v.43 no.5
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    • pp.742-749
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    • 2016
  • Boar taint, an unpleasant odor of pork, is associated with two substances including androstenone and skatole. Testosterone is a steroid hormone as well as a strong predictor for androstenone secretion. Isoflavones of soy origin play a role in modulating the metabolism of sex hormones. Although several methods responsible for reducing boar taint are under investigation, the precise mechanism by which isoflavones reduce testosterone has not yet been identified. The objective of the current study was to investigate the effects of isoflavones extracted from a soy by-product on the concentration of serum testosterone in mouse. A total of 24 mice were supplemented with basal diet (control), daidzin plus genistin mix (T1), or isoflavone extracts (T2). After 11 days of treatment, size and weight of testis, as well as the concentration of sex hormones, including testosterone and estrogen, were analyzed. There was no difference in size or weight of testis from mice among control, T1, and T2. Serum testosterone levels were significantly decreased (p < 0.05) both in T1 and T2 when compared with the control group. Furthermore, estrogen concentration in blood was increased (p < 0.05) in T2 (numerically increased in T1) compared with the control group. Taken together, the use of isoflavones extracted from soy by-products would be a plausible strategy for reducing testosterone level, ultimately reducing boar taint without castration of piglets.

Evaluation of Antifreeze Proteins on Miniature Pig Sperm Viability, DNA Damage, and Acrosome Status during Cryopreservation

  • Kim, Daeyoung
    • Journal of Embryo Transfer
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    • v.31 no.4
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    • pp.355-365
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    • 2016
  • The cryopreservation of sperm has become the subject of research for successful artificial insemination technologies. Antifreeze proteins (AFPs), one of the factors necessary for effective cryopreservation, are derived from certain Antarctic organisms. These proteins decrease the freezing point of water within these organisms to below the temperature of the surrounding seawater to protect the organism from cold shock. Accordingly, a recent study found that AFPs can increase the motility and viability of spermatozoa during cryopreservation. To evaluate this relationship, we performed cryopreservation of boar sperm with AFPs produced in the Arctic yeast Leucosporidium sp. AFP expression system at four concentrations (0, 0.01, 0.1, and $1{\mu}g/ml$) and evaluated motility using computer assisted sperm analysis. DNA damage to boar spermatozoa was measured by the comet assay, and sperm membrane integrity and acrosome integrity were evaluated by flow cytometry. The results showed that motility was positively affected by the addition of AFP at each concentration except $1{\mu}g/ml$ (p<0.001). Although cryopreservation with AFP decreased the viability of the boar sperm using, the tail DNA analyses showed that there was no significant difference between the control and the addition of 0.1 or $0.01{\mu}g/ml$ AFP. In addition, the percentage of live sperm with intact acrosomes showed the least significant difference between the control and $0.1{\mu}g/ml$ AFP (p<0.05), but increased with $1{\mu}g/ml$ AFP (p<0.001). Our results indicate that the addition of AFP during boar sperm cryopreservation can improve viability and acrosome integrity after thawing.

Effects of Discontinuous Percoll Gradient Containing Alpha-linolenic Acid on Characteristics of Frozen-thawed Boar Spermatozoa

  • Kim, Doo-San;Hwangbo, Yong;Cheong, Hee-Tae;Park, Choon-Keun
    • Journal of Animal Reproduction and Biotechnology
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    • v.35 no.1
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    • pp.58-64
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    • 2020
  • This present study was conducted to investigate protective effect of discontinuous Percoll gradient containing alpha-linolenic acid (ALA) before freezing process on viability, acrosome damage, mitochondrial activity, and oxidative stress of frozen-thawed boar spermatozoa. The separation of spermatozoa by discontinuous Percoll gradient was performed by different concentration of Percoll solution (45/90%) containing ALA combined with bovine serum albumin (BSA), and collected sperm in each Percoll layer was cryopreserved. To evaluate viability, acrosome damage, mitochondrial activity, and reactive oxygen species (ROS) level of frozen-thawed sperm, flow cytometry was used. Morphological abnormalities were observed under light microscope. In results, viability of sperm from 90% Percoll layer was higher than control and 45% Percoll group (p < 0.05). Separated sperm in 90% Percoll layer had lower acrosome damage and morphological abnormalities than control as well as viability, whereas 45% Percoll group was higher (p < 0.05). Similar with acrosome damage and abnormalities, mitochondrial activity was slightly enhanced and the population of live sperm with high ROS level was decreased by 90% Percoll separation, however, there was no significant difference. Supplementation of 3 ng/mL ALA into Percoll solution increased sperm viability and decreased population of live sperm with high ROS compared to control (p < 0.05). In conclusion, discontinuous Percoll gradient before freezing process could improve efficiency of cryopreservation of boar sperm through selection of sperm with high freezing resistance, and supplement of ALA during Percoll gradient might contribute suppression of ROS generation via stabilizing of plasma membrane during cryopreservation.

Effects of Castration and Ovariectomy on Plasma Hormone Concentration in rigs (돼지의 거세와 난소 적출이 혈중 호르몬 농도의 변화에 미치는 영향)

  • 박종대;황보종;박준철;박무균;최선호;박창식;윤종택
    • Korean Journal of Animal Reproduction
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    • v.27 no.1
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    • pp.45-51
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    • 2003
  • This study was conducted to investigate the effects of castration and ovariectomy on growth performance and plasma hormone concentration in pigs. A total of 48 pigs of 35 days of age were used. The results obtained in the present study are summarized as follows: 1. No significant difference was found in average daily gain between ovariectomy group (898.6g) and control gilt group (862.7g), and between castration group (926.0g) and control boar group (945.5g), respectively. Average daily gain of control boar group, however, was significantly higher than that of control gilt group (p<0.05). There was no significant difference in feed/gain between ovariectomy and control gilt group and between castration and control boar group, respectively. Backfat thickness was significantly (p<0.05) higher in ovariectomy or castration group than in control gilt or boar group, respectively. 2. Plasma concentration of IGF-I was significantly (p<0.05) increased during the period of 5 weeks of age (45.1 $\pm$0.72 ng/ml) to 15 weeks of age (356.3$\pm$3.05 ng/ml), and maintained constantly afterwards in control gilt group, as was in control boar group. That of ICF-I tended to be lower in ovariectomy or castration group than in control gilt or boar group, respectively. Regarding steroid hormones of estradiol-17$\beta$, progesterone, and testosterone, the concentration was extremely low at 5 weeks of age, however, increased from 11 weeks to 23 weeks of age in control gilt or boar group, while it was nearly under detection limit in ovariectomy or castration group. 3. Chemical compositions of pork loins were not affected by ovariectomy or castration, except that crude ash content was significantly (p<0.05) higher in castration group than in control boar group. These results indicated that ovariectomy or castration had no effects on growth performance and feed utilization. However, the concentration of sex steroid hormones was under detection limit in ovariectomy and castration group. Further studies, however, are needed to develope the techniques which minimize the stress related with castration or ovariectomy for the production of high quality pork.

Effects of Dietary Supplementation of Whole Garlic Powder on Semen Characteristics and Blood Antioxidant Level in Boars (수퇘지의 정액 성상과 혈중 항산화물 농도에 대한 통마늘분말 급여의 영향)

  • Park, Jun-Cheol;Cho, Kyu-Ho;Ryu, Jae-Weon;Hong, Joon-Ki;Lee, Sung-Dae;Sa, Soo-Jin;Kim, In-Cheul;Lee, Sang-Cheul
    • Journal of Animal Science and Technology
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    • v.52 no.5
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    • pp.407-412
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    • 2010
  • The objective of this study was to determine the effects of dietary supplementation of whole garlic powder (WGP) on semen characteristics and blood antioxidant level in boars. For this study, nine Duroc boars of 12 months age were used. Semen and blood samples were collected for 13 weeks, once in each week. The boars were fed the basal diet (BD; control) or BD supplemented with 3% WGP. There were no significant differences in the semen volume and sperm concentration between control and WGP group on all collection weeks. However, total sperm number per ejaculate was higher in the WGP group than that in the control group on collection weeks 6, 7 and 8 (P<0.05). Also, on collection weeks 5, 6, 7 and 8, mean of total ejaculated sperm numbers per boar were significantly higher in the WGP group compared to control group (P<0.05). On the other hand, ejaculation frequency per boar (boar's libido) and total ejaculated sperm number per boar were significantly increased in the WGP group compared to the control group, respectively (P<0.05). Although there was no difference in polyphenol level in seminal plasma between two treatment groups, polyphenol level in blood serum was significantly higher in the WGP group on collection weeks 9, 12 and 13 (P<0.05). These results indicate that dietary supplementation of 3% WGP improves boar libido and semen productivity such as ejaculation frequency per boar, total sperm number per ejaculate, mean of total ejaculated sperm number per head, and elevate the blood level of antioxidant (polyphenol) in boar serum.