• Title/Summary/Keyword: Liquid Semen

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Antibiotic Resistant Microbial Contamination (Enterobacter cloacae) Derived from Egg Yolk and Frozen Semen Extender in Porcine In Vitro Fertilized Embryos

  • Kwak, Seong-Song;Jeong, Se-Heon;Jang, Seung-Hoon;Jeon, Yu-Byeol;Nam, Young-Hee;Biswas, Dibyendu;Lee, Wan-Kyu;Hyun, Sang-Hwan
    • Journal of Embryo Transfer
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    • v.25 no.4
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    • pp.267-272
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    • 2010
  • The present study was to investigate the source of contamination during semen processing for in vitro uses. In the present study, frozen semen was prepared from liquid semen in our laboratory for in vitro fertilization (IVF) experiments due to lack of fresh semen. Antibiotics were added in the frozen semen extender (kanamycin and gentamicin) and in vitro culture (IVC) medium (gentamicin) for further inhibiting growth of microorganisms. Nevertheless, proliferations of microorganisms were observed in IVC culture drop during culturing of IVF embryos using frozen semen. Randomly 3 samples were taken from the liquid semen, frozen semen and egg yolk. Contaminated IVC medium, frozen-thawed semen, liquid semen and egg yolk were cultured in de Man, Rogosa and Sharpe (MRS) agar medium. Whitish colonies were detected in contaminated IVC drop, frozen-thawed semen samples and egg yolk but no colonies were formed in liquid semen samples. Gram-negative and rod-shaped identical bacteria were found in both frozen-thawed semen sample and contaminated IVC drop and egg yolk samples. Enterobacter cloacae were confirmed by API 20E kit according to manufacturer's instruction with identification value (% ID) 94.3% and T index 0.88. Antibiotic susceptibility tests were done according to Clinical and Laboratory Standards Institute (CLSI) by using ampicillin, amikacin, cephalothin, gentamicin, kanamycin, tetracycline, oxytetracycline, sulfamethoxazole trimethoprim, norfloxacin and ciprofloxacin test. Among them Enterobacter cloacae were resistant to ampicillin, amikacin, cephalothin, gentamicin, kanamycin but susceptible to tetracycline, oxytetracycline, sulfamethoxazole trimethoprim, norfloxacin and ciprofloxacin. From these findings it could be suggested that this contamination sources might be from egg yolk.

Effects of Sperm Number and Semen Type on Sow Reproductive Performance in Subtropical Area

  • Kuo, Y.H.;Hnang, S.Y.;Lee, K.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.1
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    • pp.6-9
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    • 2000
  • The purpose of this study was to evaluate the effect of lower numbers of sperm $(3{\times}10^9)$ per dose liquid semen and type of semen used in artificial insemination (AI) on sow reproductive performance in subtropical area. Semen was supplied by two commercial AI centers. A total of 671 female pigs from seven farms were inseminated with either $3{\times}10^9$ or $5{\times}10^9$ sperm per dose. Two types of semen were used: heterospermic semen from two boars of the same breed and homospermic semen from a single boar. After insemination, conception rate, farrowing rate, total litter size, and number of dead piglets were recorded. The analysis of variance indicated that there was no significant effect of interactions between pig farm, type of semen, or number of sperm on any of the traits measured. There were significant differences in conception rate, farrowing rate, and total litter size among pig farms (p<0.05). The effect of number of sperm per dose liquid semen ($3{\times}10^9$ or $5{\times}10^9$) was not significant. Sows inseminated with homospermic semen showed significantly higher conception and farrowing rates but significantly lower total litter size (p<0.05). In conclusion, the number of sperm per dose liquid semen for AI could be lowered to $3{\times}10^9 $ without affecting reproductive performance in subtropical areas like Taiwan.

THE COST OF SEMEN PRODUCTION AND THE RATE OF CONCEPTION FOR ARTIFICIAL INSEMINATION IN CATTLE

  • Alam, J.;Akteruzzaman, M.;Rahman, A.;Sayeed, M.A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.6 no.1
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    • pp.13-17
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    • 1993
  • This study was conducted to determine the cost of production of semen and the rate of conception for artificial insemination in eight randomly selected districts of Bangladesh. A sample of 640 adopting farms were investigated. Results indicate that the cost of liquid semen per dose on full-cost and cash cast basis were Tk. 16.17 and Tk. 14.48, while the cost of locally produced exotic semen and imported semen were Tk. 31.25 and Tk. 110.00 respectively. The cost of liquid semen per insemination, per conception and per progeny on full-cost were Tk. 26.65, Tk. 50.64, and Tk. 56.27 respectively and on cash cost were Tk. 23.88, Tk. 45.37 and Tk. 50.41 respectively. The average cost of semen paid by the farmers was Tk. 14.00 and it was higher in urban areas than in rural areas. Out of the cost of A.I. centres, government had subsidized 92.16 percent and the rest 7.84 percent accrued as returns. About 40 percent of the produced semen was lost in the system which could not be used at all. The conception rate on first insemination was 53.6 percent. The rate increased to 73.2 percent upto the last insemination. The difference in conception rate between liquid semen (69.97%) and frozen semen (70.48%) was not statistically significant.

Comparison of Motility, Acrosome, Viability and ATP of Boar Sperm with or without Cold Shock Resistance in Liquid Semen at 17℃ and 4℃, and Frozen-thawed Semen

  • Yi, Y.J.;Li, Z.H.;Kim, E.S.;Song, E.S.;Kim, H.B.;Cong, P.Q.;Lee, J.M.;Park, Chang-Sik
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.2
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    • pp.190-197
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    • 2008
  • This study was designed to analyze boar sperm to compare motility, acrosome morphology, viability and ATP by various preservation methods between Duroc boar A with cold shock resistance sperm and Duroc boar B with cold shock sensitivity sperm. Semen volume, sperm concentration, motility and normal acrosome between Duroc boar A and B did not show any differences within 2 h after collection. There were no differences in sperm motility and normal acrosome between boar A and B at 1 day of preservation at $17^{\circ}C$ and $4^{\circ}C$, respectively. However, sperm motility and normal acrosome from 2 day of preservation at $17^{\circ}C$ and $4^{\circ}C$, respectively, were higher for boar A than boar B. The frozen-thawed sperm motility and normal acrosome were higher for boar A than boar B. The sperm viability and ATP concentration according to storage period of liquid semen at $17^{\circ}C$ and $4^{\circ}C$ were higher for boar A than boar B. Also, the sperm viability and ATP concentration of frozen-thawed semen were higher for boar A than boar B. In conclusion, we found out that the original quality of boar semen with cold shock resistance sperm played an important role.

Development of Intrauterine Insemination Technique in Pig (돼지의 자궁내 인공수정기술개발에 관한 연구)

  • 공일근;정금택;이정우;정수룡;오인석;유대중;이효상;김기수;배인휴
    • Journal of Embryo Transfer
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    • v.17 no.1
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    • pp.7-12
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    • 2002
  • This study was carried out to investigate the possibility of porcine artificial insemination (A·I) on fertilizing capacity using intrauterine inseminator (IUI) method and conventional A·I (CAI) method. Number of sows used in this study was 15 far IUI and 59 fur (CAI), respectively. The results obtained are as fellows: 1 . The frozen and liquid semen used for A·I showed the higher farrowing rate in liquid semen (86.4%) than frozen semen (67%). Number of pigs born per semen type showed the higher values of number of piglets with no statistical significance using frozen semen (9.7) than liquid semen (9.3). 2. The farrowing rate per parity was highest in the 3∼5th parities (100%), f311owe4 by 0∼ 2th parities (60%), and was the smallest in 6 ∼ 10th parities (25%). Number of pigs born per litter was highest in 0∼2th parities (11.3), followed by 3 ∼ 5th parities (9.2) and lowest in 6∼ 10th parities. In the number of pigs bort per litter, the sow s in the high parities delivered lower number of piglets than those in low parities with no significant difference. These results indicated that fertilizing capacity could be improved by using IUI method.

Effect of Rubus coreanus leaf and stem extract on boar spermatozoa

  • Yi, Young-Joo;Cho, Min;Heo, Jung Min;Lee, Sang-Myeong
    • Journal of Biomedical and Translational Research
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    • v.18 no.2
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    • pp.50-55
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    • 2017
  • Rubus coreanus is known to have diverse biological properties, such as free radical scavenging activity and anti-bacterial activity. In the present study, Rubus coreanus leaf and stem extract (RLSE) was used in boar semen preservation whether it has a beneficial effect on assisted reproductive technology (ART) in mammals. Boar spermatozoa were preserved in Beltsville thawing solution (BTS) in the presence of varying concentrations of RLSE ($0-10{\mu}g/mL$). Sperm motility, sperm viability, and intracellular reactive oxygen species (ROS) levels were examined after 2 days of preservation. The percentage of total motile spermatozoa and progressive motile spermatozoa improved in the spermatozoa preserved with $0.5{\mu}g/mL$ RLSE. Higher proportions of viable spermatozoa were seen in the presence of 0.5 and $1{\mu}g/mL$ RLSE than in the control. Intracellular ROS levels decreased when the spermatozoa were preserved in BTS with $0.1-1{\mu}g/mL$ RLSE. In order to examine the bacterial growth, E. coli was added to liquid semen diluted with antibiotics-free BTS in the presence or absence of RLSE. No anti-bacterial activity of RLSE against E. coli was observed during liquid semen preservation. Although there was no inhibition of E. coli growth, the addition of RLSE might help improve sperm motility and viability during boar semen preservation, suggesting it as a potential reagent for ART in mammals.

Antioxidant Supplementation Enhances the Porcine Semen Preservation Capacity

  • Chung, Ki-Hwa;Kim, In-Cheul;Son, Jung-Ho
    • Reproductive and Developmental Biology
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    • v.39 no.1
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    • pp.7-11
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    • 2015
  • Preservation of liquid semen is an important factor for breeding management in swine industry. Oxidative stress of spermatozoa during liquid preservation has a detrimental effect on sperm quality and decreases fertility. Objective of this study was to determine the effect of antioxidant, Quercetin, on capability of porcine liquid semen preservation. Freshly collected porcine semen from boars (n=3), having proven fertility was counted, diluted to $3{\times}10^7/mL$ and divided into 5 different semen extenders. Aliquots of diluted semen with different extenders were subjected to measure the pH, motility, viability and sperm DNA structure status on elapse time after preservation for 10 days. For the first 3 days, semen preserved in all 5 different extenders maintained their initial pH and either gradually decreased or increased thereafter, indicating lipid peroxidation has started. Sperm motility (r=0.52, p=0.01) and viability (r=0.55, p=0.03) had positive correlation with semen pH. Sperm motility was maintained well (p<0.05) in especially 2 extenders containing Tris and antioxidant compared to other extenders, suggesting both Tris and antioxidant worked as pH regulator and had beneficial effects on sperm characteristic during preservation. Sperm DNA structure status accessed by sperm chromatin structure assay on elapsed time after preservation, tended to be higher in semen preserved without antioxidant. Taken together, addition of antioxidant to extender prevents the sperm from oxidative stress during storage in mechanism by which antioxidant slows the lipid peroxidation, and thus reduced the reactive oxygen species in preserved porcine semen resulted in maintaining semen pH, sperm motility and viability for 7~10 days.

An Analysis of Survey Data on South Korea Boar Stud Practices (한국의 돼지 인공수정센터 현황 분석)

  • Sa, Soo-Jin;Woo, Jae-Seok;Hong, Joon-Ki;Kim, Ki-Hyun;Kim, Du-Wan;Kim, Yong-Min;Park, Hyun-Sik;Kim, Si-Joo;Chung, Ki-Hwa;Cho, Eun-Seok
    • Journal of Embryo Transfer
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    • v.30 no.3
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    • pp.129-135
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    • 2015
  • The aim of this study is to examine current status of swine AI and boar stud in South Korea using survey and data analysis. This survey included 48 boar studs registered as 'semen processing business'. The survey data were collected by direct visitation, FAX and/or telephone conversation for 7 months from June through December in 2013. 48 boar studs owned a total of 3,537 boars and the Duroc breed accounted for the highest rate (75.3%) of all boar breeds. In case of ownership, agricultural management corporations was the highest (50.0%) and followed by individual ownership (33.3%). Large-scale boar studs in terms of own over 151 boar were surveyed as 4.2% and most boar studs owned less than 100 boars (77.1%). The amount of liquid semen provided by 48 boar studs were 1,889,000 doses and each boar stud provided average of 39,000 does, which is represented for 90% consumption by sows in South Korea.

Effects of Bacterial Contamination of Extended Boar Semen Preservation Periods on Embryo Production In Vitro (돼지 액상 정액의 보관일수에 따른 오염 정도가 체외 수정란 생산 효율에 미치는 영향)

  • Kim, Y.S.;Lee, H.T.;Kim, I.C.;Ryu, J.W.;Kim, C.W.;Chung, K.H.
    • Journal of Embryo Transfer
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    • v.21 no.4
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    • pp.345-351
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    • 2006
  • The objective of this study was to investigate the effects of preservation period of porcine liquid semen on bacterial contamination and in vitro production of embryo. Extended liquid semen was prepared by three mixture of boar's ejaculates from each farm without antibiotics, and were kept in $17^{\circ}C$ semen preservation incubator until use. Sperm motility was significantly (p<0.05) decreased as semen preservation time goes by (78.7$\pm$2.4% for 1 day vs. 71.1$\pm$2.4 and 64.8$\pm$2.4% for 3 and 5 days of presentation, respectively). Quantitative of bacteria in semen was significantly (p<0.05) higher in 5 days ($57.8\pm105.2\times10^4$ Cfu) compared to 0 and 3 days ($32.1\pm76.8\times10^4$ and $26.9\pm46.6\times10^4$ Cfu, respectively) of preservation. In terms of development of in vitro fertilization of porcine embryos inseminated by preserved semen, the rate of normal fertilization (2PN) was significantly (p<0.05) decreased in 5 days (56.0$\pm$2.6%) compared to 1 and 3 days (66.0$\pm$2.7 and 64.0$\pm$2.7%, respectively) of preservation. Cleavage rate was also significantly (p<0.05) affected by preservation period (75.0$\pm$4% for 1 day, 70.0$\pm$0.3 and 71.0$\pm$0.3% for 3 and 5 days, respectively). The in vitro developmental rate of blastocyst stage embryo was significantly (p<0.05) affected by semen preservation period (15.0$\pm$1.0% for 1 day vs. 11.0$\pm$0.9 and 8.0$\pm$0.9% for 3 and 5 days, respectively). It is concluded that more than 3 days of liquid semen preservation without antibiotics increased the quantity of bacteria resulted in detrimental effect on sperm motility and decreased both normal insemination rate and the developmental rate of blastocyst stage embryo.

Comparison of Viability, ATP and In vitro Fertilization of Boar Sperm Stored at 4℃ in the Three Different Diluents

  • Yi, Y.J.;Li, Z.H.;Kim, E.S.;Song, E.S.;Cong, P.Q.;Zhang, Y.H.;Lee, S.H.;Lee, J.W.;Park, Chang-Sik
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.8
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    • pp.1127-1133
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    • 2008
  • This study was designed to develop a method of liquid storage of boar sperm at $4^{\circ}C$ by using the modified Beltsville F5 (BF5) diluent with bovine serum albumin (BSA) and N-acetyl-D-glucosamine. Boar sperm were stored in lactose-egg yolk and N-acetyl-D-glucosamine (LEN), BF5 and Golden-Pig liquid 4 (GPL4) diluents at $4^{\circ}C$ for 5 days and were examined for sperm viability, adenosine triphosphate (ATP) and in vitro fertilization (IVF). The percentage of sperm viability in GPL4 diluent was higher than in LEN and BF5 diluent from 1 to 5 days of storage at $4^{\circ}C$. The percentage of sperm viability steadily declined from 1 to 5 days of storage in the three different diluents. Sperm ATP in GPL4 diluent was higher than in LEN and BF5 diluents from 1 to 5 days of storage. Sperm ATP rapidly declined after 5 days of storage in the three different diluents. Porcine oocytes matured in vitro were inseminated with different sperm concentrations of liquid semen stored for 3 days in GPL4 diluent. The percentage of monospermic oocytes did not show any differences from 2.5 to $20{\times}10^5$ sperm/ml. However, the percentage of polyspermic oocytes in the sperm concentration of $2.5{\times}10^5$ sperm/ml was lower than in concentrations of 5, 10 and $20{\times}10^5$5 sperm/ml. The percentage of blastocysts from the cleaved oocytes at $2.5{\times}10^5/ml$ sperm concentration was significantly lower than at 5, 10 and $20{\times}10^5sperm/ml$ concentrations. In conclusion, GPL4 diluent can be stored at $4^{\circ}C$ for 5 days and showed higher sperm viability and sperm ATP concentration compared with LEN and BF5 diluents. Also, we found that GPL4 diluent can be used for IVF of porcine oocytes.