• 제목/요약/키워드: Extended semen

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Estimates of Genetic Correlations between Production and Semen Traits in Boar

  • Oh, S.H.;See, M.T.;Long, T.E.;Galvin, J.M.
    • Asian-Australasian Journal of Animal Sciences
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    • 제19권2호
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    • pp.160-164
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    • 2006
  • Currently, boars selected for commercial use as AI sires are evaluated on grow-finish performance and carcass characteristics. If AI sires were also evaluated and selected on semen production, it may be possible to reduce the number of boars required to service sows, thereby improving the productivity and profitability of the boar stud. The objective of this study was to estimate genetic correlations between production and semen traits in the boar: average daily gain (ADG), backfat thickness (BF) and muscle depth (MD) as production traits, and total sperm cells (TSC), total concentration (TC), volume collected (SV), number of extended doses (ND), and acceptance rate of ejaculates (AR) as semen traits. Semen collection records and performance data for 843 boars and two generations of pedigree data were provided by Smithfield Premium Genetics. Backfat thickness and MD were measured by real-time ultrasound. Genetic parameters were estimated from five four-trait and one five-trait animal models using MTDFREML. Average heritability estimates were 0.39 for ADG, 0.32 for BF, 0.15 for MD, and repeatability estimates were 0.38 for SV, 0.37 for TSC, 0.09 for TC, 0.39 for ND, and 0.16 for AR. Semen traits showed a strong negative genetic correlation with MD and positive genetic correlation with BF. Genetic correlations between semen traits and ADG were low. Therefore, current AI boar selection practices may be having a detrimental effect on semen production.

Evaluation of Boar Sperm Viability by MTT Reduction Assay in Beltsville Thawing Solution Extender

  • Byuna, J.W.;Choo, S.H.;Kim, H.H.;Kim, Y.J.;Hwang, Y.J.;Kim, D.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • 제21권4호
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    • pp.494-498
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    • 2008
  • MTT (3-(4, 5-dimethyl thiazol-2-yl)-2, 5-diphenyl tetrazolium bromide) reduction assay is a method that validates the viability of an active cell. Dehydrogenase in mitochondria converts yellow colored insoluble tetrazolium salt to purple colored water-soluble formazan. Sperm also have mitochondria in the midpiece, therefore sperm viability could be evaluated by MTT reduction assay. Several studies have already demonstrated the capability of application of the MTT reduction assay to sperm of several species in Hepes-BSA buffer. Because most liquid semen was diluted in extender like BTS (Beltsville Thawing Solution), Modena or Androhep when it is used or transferred, semen needed another dilution in Hepes-BSA buffer to assess sperm viability. In this study, we evaluated boar sperm viability especially in BTS extended semen and compared the efficiency of this test with eosin-nigrosin staining. We used the fresh BTS extended semen from a local A.I center. Semen sample was diluted to $3.0{\times}10^7$ sperms/ml in BTS. The rates of formazan production were measured in 96-well microtiter plates immediately and 1h after incubation at $17^{\circ}C$ using a spectrophotometer at wave length 560 nm. Simultaneously, split samples of the same semen were tested, using eosin-nigrosin staining to compare the efficiency of the MTT assay of sperm viability in BTS. The correlation between the results of these tests was calculated using Student-t test and ANOVA. The results revealed a strong correlation between the results of MTT reduction rate and the results that were simultaneously determined by eosin-nigrosin staining at 1 h. In conclusion, the MTT reduction test was an effective and simple method to validate sperm viability and it could be used as a simple tool to evaluate sperm viability in the local A.I center and laboratory.

Potential of watermelon (Citrullus lanatus) to maintain oxidative stability of rooster semen for artificial insemination

  • Jimoh, Olatunji Abubakar;Akinola, Micheal Olawale;Oyeyemi, Bolaji Fatai;Oyeyemi, Wahab Adekunle;Ayodele, Simeon Olugbenga;Omoniyi, Idowu Samuel;Okin-Aminu, Hafsat Ololade
    • Journal of Animal Science and Technology
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    • 제63권1호
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    • pp.46-57
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    • 2021
  • Fruits with antioxidant enrichment can be an economically affordable supplement for mitigating oxidative damage prone spermatozoa membrane pathologies. Computer-assisted sperm analyzer and oxidative status were utilized to evaluate the impact of watermelon (Citrullus lanatus) fortification of dextrose saline as diluent for rooster semen and fertility response of hens inseminated. Watermelon juice and dextrose saline were used to formulate diluent of 7 treatments consisting of unextended semen (positive control), 10%, 20%, 30%, 40%, 50% and only dextrose saline (negative control) designated as Treatments 1-7. Pooled semen was obtained from fertile roosters and equilibrated with diluents at ratio 1:2 in the various treatments and were evaluated using computer software coupled microscope and seminal oxidative status assay. 168 laying hens randomly divided into 7 treatment of 8 replicates and 3 hen per replicate. Hen were everted, and semen (2 × 108 Spermatozoa) deposited intra-vagina and eggs collected over 8 weeks to assess fertility and hatchability of eggs laid. The result obtained revealed that watermelon-dextrose saline rooster semen diluent enhanced progressive motility, sperm kinetics and lowered non-progressive motility in T2-T6 compared to T7 over the 3 hours of evaluation. Watermelon addition to rooster semen diluent enhance the antioxidant capacity of rooster semen and lowered lipid peroxide generation. The percentage fertility was highest in T3 (81.01%) and T4 (81.24%) with lowest value obtained in T7 (73.46%). The hatchability of eggs set of hens inseminated with undiluted semen (71.46%) was lower than values for hens inseminated with watermelon inclusive extended semen (75.71%-80.39%). The optimal inclusion of 30%-40% watermelon in dextrose saline diluent enhance rooster semen kinetics, seminal oxidative stability and egg fertility.

Effects of Different Concentrations of Escherichia coli and Days of Preservation on Boar Sperm Quality

  • Chung, Ki-Hwa;Kim, In-Cheul;Son, Jung-Ho
    • Reproductive and Developmental Biology
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    • 제37권4호
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    • pp.213-217
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    • 2013
  • The objective of this study was to investigate the effect of bacterial contamination on elapsed time after preservation on boar semen. Known numbers of Escherichia coli (E. coli) were inoculated to freshly ejaculated semen and sperm parameters such as viability, motility, agglutination, acrosome integrity and hypo-osmotic swelling test were performed during 7 days of liquid preservation. Semen samples were prepared using antibiotic free BTS extender and 4 different levels of E. coli were treated to semen with following concentrations; 3,000, 5,000, 7,000, 10,000 CFU/ml of sperms. Semen samples were preserved at $17^{\circ}C$ for 7 days in semen storage until analyzed. Aliquots were subjected to measure the sperm viability, motility and agglutination using computer assisted sperm analysis (CASA) system, acrosome integrity was performed using chlortetracycline (CTC) staining method and hypo-osmotic swelling test was performed using hypotonic solution from day 1 (day of semen collection) to 7. Detrimental effects on sperm motility and viability were observed 3 days after preservation at the level of 5,000 CFU/ml (p<0.05). Percentage of sperm abnormality was higher (p<0.05) in over 5,000 CFU/ml groups. Sperm agglutination rate was also significantly higher (p<0.05) in groups of 5,000 and 7,000 CFU/ml. The rate of acrosome reacted sperm was higher as preservation time goes in all the samples but the pattern was clearly higher among E. coli contaminated groups (p<0.05). The sperm membrane integrity in terms of hypo-osmotic test, E. coli affects little compared to other sperm parameters. The deleterious effects observed due to the bacterial contamination in semen suggest that importance of hygiene protocol to minimize the bacterial contamination during semen collection and processing.

Thoroughbred 정액의 액상 보존에 관한 연구 (Preservation of Extended Thoroughbred Semen at Low Temperature)

  • 고태혁;김한섭;이상호;송해범
    • 한국가축번식학회지
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    • 제14권3호
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    • pp.199-204
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    • 1990
  • Equine semen was analysed for its survival after storage under different conditions. Total 12 ejaculates from 2 Thoroughbred were analysed for general characterisitcs and preservation at low temperature. The sperm fraction, concentration, nd the rate of motile spermatozoa were 57.91ml per ejaculate, 2.18$\times$108/ml and 74.1%, respectively. The survival rate of spermatozoa was highest when diluted semen with E-Z Mixin was stored at 7~8$^{\circ}C$. The optimum survival rate(>54%) can be obtained upto 24h at 7~8$^{\circ}C$. However only 10% spermatozoa survived after 5h storage at 7~8$^{\circ}C$ without use of E-Z Mixin. Other ranges of temperature(15$^{\circ}C$ and room temperature) gave less survival rates(<25%). These results indicate that the extender could be used as a basic solution for the preservation of equine spermatozoa at low temperature. It also provides a practical method for short-term storage of collected equine semen in a simple manner.

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소혈청알부민과 당류가 돼지 동결정자의 생존성 및 두모형태에 미치는 영향 (Effects of Bovine Serum Albumin and Sugars on Sperm Livability and Acrosome Morphology of Frozen-thawed Boar Semen)

  • 윤종택;임경순;이용빈
    • 한국가축번식학회지
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    • 제10권1호
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    • pp.19-26
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    • 1986
  • This experiment was carried out to investigate the effect of bovine serum ablumin (BSA), sugars, glycerol equilibration time, straw size and thawing method on the survival index and the morphology of frozen boar spermatozoa. The results obtained were summarized as follow: 1. When the semen frozen in BF5 dilutor as pellet form was thawed in BTS at 37$^{\circ}$and 50$^{\circ}C$, BF5 dilutor with fructose showed higher sperm survival index than that with dextrose, however, when the semen was thawed on dry test tube at 37$^{\circ}C$, BF5 dilutor with sucrose showed higher sperm survival index than with other sugars. 2 When the semen forzen in BF5 dilutor with straw and thawed at 37$^{\circ}C$, BF5 dilutor with dextrose showed higher sperm survival index than those with other sugars, and there was no difference in sperm survival index between 0.5 and 1.0 ml straws. 3. The sperm survival index of frozen sperm was significantly (P<0.05) improved due to addition of BSA (0.05%) to BF5 dilutor. 4. When the extended semen with BF5 dilutor contatining 0.01 to 0.05% of BSA was frozen in the straw, the semen without glycerol equilibration showed significantly (P<0.05) higher sperm survival index than those with 2, 4 and 6 hrs glycerol equilibration time. 5. The sperm frozen in BF5 dilutor with dextrose or fructose, sucrose and raffinose showed 77 to 88% in normal acrosome rate and no difference among sugars. 6. The frozen semen showed lower normal acrosome rate than the first and second diluted semen, whereas the frozen semen showed higher swollen, damaged and missing acrosome rate than the first and second diluted semen. 7. Damaged and missing acrosome rate of sperm head due to freezing was somewhat inhibited by addition of BSA (0.01 to 0.05) to the BF5 dilutor.

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Assessment of Sperm Characteristics in Fresh and Frozen Semen of Miniature-Pig

  • Lee S. H.;Kim T. S.;Cheong H. T.;Yang B. K.;Kim C. I.;Park C. K.
    • Reproductive and Developmental Biology
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    • 제28권4호
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    • pp.261-265
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    • 2004
  • The present study was conducted to assess sperm characteristics in miniature-pig. The semen samples were transported to the laboratory at 17℃ within 3 hours after collection. The extended semen was stored at 17℃, and sperm quality was evaluated at 0, 1, 3, 5 and 7 days after storage. The semen volume of miniature-pig (62±22㎖) was significantly (p<0.05) lower than that of Duroc (155±25㎖) and Yorkshire (154±23㎖). Significant differences were also observed in sperm concentrations. During 3 days of storage, sperm viability did not differ among miniature-pig, Duroc and Yorkshire. However, the viability was significantly (p<0.05) lower in miniature-pig than in Duroc and Yorkshire semen after Day 3 of storage. In abnormality, acrosome intactness and intensity, there were no differences among miniature-pig, Duroc and Yorkshire semen. On the other hand, the viability of frozen-thawed sperm in miniature-pig was significantly (p<0.05) lower than in that of Duroc and Yorkshire. This study also examined CTC patterns in frozen-thawed spermatozoa. The rates of AR pattern were higher in miniature-pig than in Duroc and Yorkshire. However, no difference was found in F, B and AR patterns. The results of present study suggest that further research is necessary to develop of semen extender and freezing methods to improve sperm quality in miniature-pig.

개 정맥의 동결 및 융해후 정자의 생존성 및 수정능획득 판정을 위한 HOS test 및 CTC test (Studies on HOSS test and CTC test for Viability and Capacitation of Frozen-thawed Canine Sperm)

  • 김용준;지동범;오홍근
    • 한국임상수의학회지
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    • 제17권2호
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    • pp.431-437
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    • 2000
  • Evaluation of viability and capacitation of canine sperm is of great importance to deter- mine good condition for freezing canine semen and consequently to improve conception rate by arti-ficial insemination. Semen were collected from nine male dots which had been proved to be fertile in the post and the semen were treaded for freezing procedure. Semen were thawed at 37$^{\circ}C$ for 30 seconds. In this study, hypoosmotic swelling(HOS) test and chlortetracycline(CTC) test were per- formed to evaluate post-thaw viability and capacitated status of sperm, respectively. In HOS test far canine sperm, the highest percentage of curled sperm was shown at 60 mOsm. In HOS test for canine semen, there were considerably significant correlation between HOS values and sperm motil- ity(r=0.9064, p<0.01) and converse correlation between HOS values and sperm abnormality(r=- 0.6905, p<0.05). The sperm viability and HOS-values for chilled extended semen were significantly decreased from 0 to 72 hours during storage at 5$^{\circ}C$ (p<0.05). Of the media added to canine semen after thawing, the most capacitated sperm were shown in CCM(p<0.01), and then This Fructose Cit- rate(TFC) medium with calcium from 3 hours after incubation with media. It was concluded that HOS test is of great value to determine the viability and motility of canine sperm, whereas CTC test is usable to determine the capacitated status. Consequently, both tests were thought to be useful as the additional tests to standard semen analysis.

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돼지 정액을 저장하는 동안 정자에 미치는 산화스트레스 (Oxidative Stress in Spermatozoa during Boar Semen Storage)

  • 이승형
    • 생명과학회지
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    • 제33권7호
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    • pp.586-592
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    • 2023
  • 돼지 정액을 저장하는 동안 산화스트레스의 발생은 정자의 질과 생존에 영향을 미치는 중요한 인자이다. 정액의 저장은 온도 변화, 동결보호제 등의 다양한 스트레스 인자에 노출되어 있다. 이러한 정자 내에서의 산화스트레스는 활성산소종의 생성에 의해 발생되며, 이는 지질, 단백질, DNA와 같은 세포를 구성하는 물질에 산화적으로 손상을 일으킨다. 활성산소종과 항산화물질의 균형있는 체계는 정자의 생존과 그 기능을 유지하는 데 중요한 역할을 한다. 정액을 장기간 보존하게 되면 활성산소종의 수준이 증가하여 정자의 운동성, 막 온전성, DNA 온전성에 영향을 미치게 된다. 또한, 활성산소종에 의해 유도된 지질과산화 반응은 정자막의 유동성과 안정성에 영향을 미쳐 정자의 운동성을 감소시킨다. 그리고, DNA의 산화적 손상은 DNA 단편화를 일으켜 정자의 DNA 온전성을 손상시킬 수 있다. 결론적으로, 정액을 보관하는 동안 발생되는 산화스트레스는 정자의 질과 기능을 유지하는 데 중요하다. 따라서, 산화스트레스의 기본적인 메커니즘과 정자의 기능에 미치는 영향을 이해하는 것은 산화스트레스로부터의 손상을 최소화하고, 효율적이고 기능적인 정자의 저장 방법을 개선하기 위한 효과적인 전략과 연구 개발을 위해 중요할 것으로 판단된다.

Study on Suitable Semen Additives Incorporation into the Extender Stored at Refrigerated Temperature

  • Bhakat, M.;Mohanty, T.K.;Raina, V.S.;Gupta, A.K.;Pankaj, P.K.;Mahapatra, R.K.;Sarkar, M.
    • Asian-Australasian Journal of Animal Sciences
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    • 제24권10호
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    • pp.1348-1357
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    • 2011
  • The objective of this study was to compare the effect of Butylated Hydroxy Toluene (BHT), Pentoxifylline (PTX) and ${\alpha}$-tocopherol (Vit E) on semen quality parameters of Karan Fries bulls. The fortification of extender by various semen additives improves motility as well as fertility of spermatozoa. Split samples of 24 ejaculates of four Karan Fries bulls were extended in extender with or without various additives such as BHT, PTX and Vit E, and performance was evaluated at an interval of 0, 24, 48 and 72 h at refrigerated temperature (4-$7^{\circ}C$). Results of the present study revealed that addition of BHT, PTX and Vit E in extender improved sperm cell function, such as motility, viability, HOST, and acrosome integrity, as compared to the control during liquid storage up to 48 h of preservation at refrigerated temperature. There was no significant (p<0.05) difference between any of the additives up to 48 h of preservation. Overall, the results showed a significant (p<0.05) deterioration in motility after each storage interval. The results showed a significant deterioration in the acrosome integrity and plasma membrane integrity up to 48 h; subsequently, there was not much degradation of both the semen quality parameters. There was a significant increase in spermatozoal tail and total abnormality after each storage interval at refrigerator temperature (4 to $7^{\circ}C$); however, the head and mid-piece abnormalities were almost unaffected. Tail and total abnormality were least in extender fortified with BHT, PTX and Vit E at different hours of incubation as compared to the control. The addition of 1.5 mM BHT, 3.6 mM PTX and 1 mg/ml Vit E in the semen extender has more beneficial effect in terms of semen quality and preservability of spermatozoa.