• 한국가금학회지
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• 제48권4호
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• pp.185-191
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• 2021

닭 정자는 저온 환경에서 생존하는 능력을 보유하고 있으나 정자의 운동성과 첨체 손상에 대한 연구는 상세히 알려져 있지 않다고 판단된다. 본 연구에서는 닭 정자를 BPSE 및 Lake 희석액으로 희석하고 보존 용기를 1.5 mL e-tube와 0.5 mL 스트로에 보존하여 정자 장수성을 조사하였다. 밀봉된 0.5 mL 스트로에 보존된 정자의 운동성이 3일동안 저온보존 시 더 높은 것으로 관찰되었다(68.6±3.1% vs. 22.1±5.7%). BPSE로 희석하여 0.5 mL 스트로에 보존된 닭 정자의 운동성 또한 Lake 희석액보다 더 높게 관찰되었다(57.7±5.6% vs. 37.7±5.4%). 첨체 온전성도 0.5 mL 스트로에 보존하는 것이 우수하였고 냉장보존일이 증가함에 따라 손상된 첨체 비율이 증가하였다. 본 연구에서 0.5 mL에 밀봉된 닭 정자는 저온 보존에 이용될 수 있으며 정자의 운동성을 높여주며 첨체 온전성을 증가시키는 것으로 관찰되었다.

• Reproductive and Developmental Biology
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• 제35권3호
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• pp.203-207
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• 2011

The objective of this study was to determine the effect of semen extenders on the motility, viability and fertility in vitro of spermatozoa during storage of fresh boar semen diluted in different commercial extenders used for pig artificial insemination (AI). In this experiment, semen were diluted in Androhep plus, Beltsville Thawing Solution (BTS), Modena, Seminark and Vitasem LD. Five ejaculates were collected from three Duroc boars and sub-samples were diluted ($30{\times}10^6$ spermatozoa/ml) in different extenders. Semen was stored at $170^{\circ}C$ for 10 days. Sperm motility and viability was assessed using Computer-Assisted Semen Analysis (CASA) and flow-cytometry on 1, 3, 5 and 10 day post collection The motility of spermatozoa stored in different extenders was gradually decreased by increasing the duration of storage of semen. However, there was not significant1y different in the sperm motility and viability among other extenders. On the other hand, the in vitro-matured oocytes were fertilized and cultured in vitro to assess the fertility of boar spermatozoa stored for 3 and 10 days in different extenders. The percentage of morula and blastocyst were taken as indicators of fertility in vitro of spermatozoa. Therefore, there were no differences in the rate of embryos developed to the molular and blastocyst stage. There were no differences in the motility and fertility in vitro among 5 kinds of commercial boar semen extenders.

• Reproductive and Developmental Biology
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• 제40권1호
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• pp.1-5
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• 2016

This study investigated the effects of L-carnitine (LC) and nicotinic acid (NA) on sperm viability during liquid storage at $18^{\circ}C$ in miniature pigs. $10{\mu}M$ LC and 30 mM NA, combined LC and NA (LN) were treated in fresh semen for 3, 7, and 10 days. In results, sperm survival increased in NA- and LN-treated semen on 7 and 10 days (p<0.05), mitochondrial integrity of live sperm increased in LN-treated semen on 7 days (p<0.05), but not NA-treated semen. In addition, we examined the acrosome reaction of sperm in miniature pigs. LC and NA did not influence on acrosome reaction of boar sperm. In conclusion, LC and NA effectively maintained the viability and quality of sperm during long-term storage in miniature pigs, suggesting that the combined LN may be useful for improving the semen extender for long-term liquid storage in pigs.

• Asian-Australasian Journal of Animal Sciences
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• 제15권5호
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• pp.633-636
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• 2002

Six adult Garole rams maintained under a semi-intensive system were used as semen donors for this study. Semen was collectied daily during the monsoon season with the help of an artificial vagina and examined for its quality characteristics. Ejaculates of thick consistency, rapid wave motion, ${\geq}80%$ motility and intense movement of motile spermatozoa were diluted at the rate of 1:1 with egg yolk McIllvaine glucose diluent at $30^{\circ}C$ in water bath. Estrus in ewes was detected by parading aproned rams of proven vigour at 12 h intervals. The ewes (54 Malpura and 23 Avikalin) in estrus were artificially inseminated with fresh diluted ram semen. The overall conception rate was 94.8%, (range 91.7 to 100%). The overall lambing percent was 80.5 with a range of 75.0 to 84.6%. There was no significant (p>0.05) difference in lambing and conception rate because of individual rams. Fertility was significantly lower (p<0.05) in ewes of less than two years and more than six years of age. Breed (Malpura and Avikalin) effect was not observed in conception and lambing rate (p>0.05). No significant difference (p>0.05) in birth weight and 12 month weight was observed between Garole${\times}$Avikalin and Garole${\times}$Malpura crossbred lambs but there was significant (p<0.05) difference at three month and six months body weight of both the crossbred lambs.

• 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.

• 한국수정란이식학회지
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• 제24권3호
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• pp.207-212
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• 2009

The aim of present experiment was to examine commercial synthetic extender(AndroMed) for semen cryopreservation of Korean Black Bull. Semen was collected from a Korean Black Bull using an artificial vagina and transported to the laboratory. The semen was diluted 1:1 by AndroMed. The pellect was diluted to final sperm concentration of $5{\times}10^5/ml$ by doubling in every 10 minutes at $4^{\circ}C$ cold chamber. The semen was equilibrated for 1 hr at cold chamber and packed to 0.5 ml straw. The semen straws were located above 5 cm of liquid nitrogen for 5 minutes, above 5 cm for 10 minutes and above 10 cm for 10 min. And then the frozen straw was plunged to $LN_2$. The presented straws were examined the viability and motility after thawed at $37^{\circ}C$ water bath. Hanwoo semen was used as KPN (Korea Proven Bull Number) in this experiment. The survival rates was significantly higher in fresh semen than frozen semen ($80{\pm}14%\;and\;43{\pm}11%$). However, the motility rates was similar (80.7% and 66.4%). The survival and motility rates were higher in 5cm, 10 min treatment group than the other two groups in straw-located height and duration above $LN_2$ ($50{\pm}14%$ and 70.7% vs, 33.18% and $65{\pm}7%$ vs, 30.14% and 65.7%, respectively). The development rates to cleavage was higher in Black Cow than Hanwoo semen (62.2%, 64.4%), However, The development rates to blastocyst was higher in Hanwoo than Black cow semen (25.9%, 23.0%). In conclusion. The present results that acceptable fertilization and cryopreservation could be obtained by in vitro fertilization with frozen-thawed semen using a synthetic semen extender (AndroMed).

• 한국수정란이식학회지
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• 제25권4호
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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.

• Animal Bioscience
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• 제37권7호
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• pp.1177-1184
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• 2024

Objective: Semen cryopreservation is an effective method of preserving genetic material, particularly in native chicken breeds facing a substantial decline. In this study, we evaluated the quality of frozen/thawed rooster semen treated with different concentrations of oral administrations of black ginger (Kaempferia parviflora: KP) extract and determined its fertility. Methods: Thirty-two Thai native roosters (Pradu Hang Dum, 42 weeks old) were used in this study. The treatments were classified into four groups according to the concentration of KP extract administered to the roosters: 0, 100, 150, and 200 mg/kg body weight. The quality of fresh semen was analyzed before cryopreservation. Post-thaw sperm quality and fertility potential were determined. Also, lipid peroxidation was determined. Results: The results showed that sperm concentration and movement increased in roosters treated with 200 mg/kg of KP extract (p<0.05). The malondialdehyde (MDA) in the roosters receiving 200 mg/kg KP extract was lower than that in the other but had an insignificant difference within the KP treatment groups (p>0.05). The highest MDA levels were observed in the control group (p<0.05). The percentage of motile sperm (total motility and progressive motility) after semen thawing was higher in roosters that received 150 and 200 mg/kg KP extract than in those that received 100 mg/kg KP extract and the control (p<0.05). MDA levels decreased significantly in roosters that received 150 and 200 mg/kg KP extract than in those that received 100 mg/kg KP extract and the control (p<0.05). Fertility and hatchability were greater in the KP150 and KP200 groups than in the KP100 and control groups (p<0.05). Conclusion: The optimal amount of KP extract influencing initial sperm quality was determined to be 200 mg/kg. However, 150 mg/kg was the optimal low dosage of KP extract administration that maintained sperm quality and fertility following semen cryopreservation.

• Reproductive and Developmental Biology
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• 제30권1호
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• pp.71-74
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• 2006

Hypoosmotic swelling test (HOST) is used for evaluating the plasma membrane function and fertilizing ability in mammal spermatozoa. However, HOS solutions and experimental conditions have not been determined clearly for assessing canine spermatozoa. This study was conducted to examine the HOS solutions and assay conditions, including incubation time (30 to 120 min), storage temperature (4, 17 and $20^{\circ}C$), semen status (fresh and frozen). Maximum spermatozoal plasma membrane swelling was obtained in an 150 mOsm Na-citrate/Fructose solutions with an incubation time for 45 min. The storage temperature and semen status affected the percentage of HOS positive spermatozoa. The HOS test adapted to canine spermatozoa in this study was simple and highly consistent assay with good repeatability. The optimal condition of HOST in canine spermatozoa is an 150 mOsm Na-citrate/Fructose solutions with an incubation time for 45 min regardless of semen storage temperature and semen status.

• 한국가금학회지
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• 제42권2호
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• pp.109-116
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• 2015

본 연구에서는 육종이 필요한 종계장에서 슬라이드 위에 도말된 닭 정액시료를 간편하게 염색하여 정자의 염색질 이상성과 운동성을 유추할 수 있는 기법을 확립하고자 실시하였다. 오계 정자 도말 슬라이드를 Diff-Quik 시약으로 염색하여 관찰하면 신선정액에서는 정자 생존성이 93.53%, 82.42% 및 90.63%일 때, Diff-Quik 염색질의 정상도는 87.96%, 74.25% 및 85.10%로 관찰되었다. 동일한 시료를 동결하고 융해후 정액의 생존성은 조사하면 69.58%, 61.98% 및 72.20%로 관찰되었으며, 염색질의 정상도는 58.91%, 48.49% 및 63.34%로 관찰되었다. 융해된 동결정액에서 활력이 우수한 정자를 쉽게 관찰하기 위하여 정자를 HS-1 희석액으로 재 희석하고, $37^{\circ}C$에서 가온하여 도말하면 염색된 정자의 두부에서 활력이 우수한 정자의 비율을 간단히 유추할 수 있음을 보여주었다. 특히, 신선 정자에서 정상 염색질을 가진 정자의 비율과 생존성이 상관관계는 매우 높은 것으로 판단되며, 동결정자에서도 상관관계가 높다고 추정되었다. 이러한 결과는 Diff-Quik 염색방법으로 닭 정액의 품질을 정자의 염색질의 이상성 유무로 판단할 수 있음을 보여주고 있다. 특히 본 연구에서 제시된 방법은 닭 정자의 우수성을 판단하는데 유용할 것으로 판단되며, 닭 정액 시료를 준비할 때 준비한 도말슬라이드를 현미경적 관찰에 의하여 활성화된 정자의 비율과 정상 염색질을 가진 정자의 비율을 추정하는 방법을 제시하였으며, 이는 인공 수정에 필요한 현장에서 수컷 개체의 종축 이용성을 쉽게 판단할 수 있는 근거를 마련할 수 있음을 의미한다.